Anti Protein Research Articles

Gas Chromatography-Mass Spectroscopy and Fourier Transform Infrared Profiling of the Bioactive Compounds Present in Methanol Leaf Extract of Solanum Aethiopicum from Imo State, Nigeria

The study aimed to access the bioactive compounds and functional groups present in Solanum aethiopicum using the Gas Chromatography coupled with mass spectrometer (GCMS) and Fourier Transform Infrared Spectroscopy (FTIR). The GCMS showed Solanum aethiopicum showed a total of thirty-seven bioactive compound with the highest abundance being Squalene (21.13%), Palmitoleic acid (16.63%), 13-Octadecenal (16.63%) and 9-Octadecenoic acid (17.62%). Fourier Transform Infrared Spectroscopy of Solanum aethiopicum showed the presence of functional groups which include alkanes, alkenes, alkynes, primary (10) and secondary (20) amines, amides, imino, epoxy, peroxides, carboxylic esters, ethers, primary (10), secondary (20) and Tertiary (30) alcohols, phenols, aldehydes, thiols, cyanate, isocyanates and alkyl halides. These bioactive compounds with reference to literature revealed a variety of biological functions spanning across antibacterial, anti-inflammatory, anti-oxidant, analgesic, antimalarial, antifungal, anti-tumour, neuroprotective, anti-tumour, anti-cancer, anti-seizure, anti-allergy, anti beta-amyloid protein aggregation.

Cohort profile: follow-up of a household cohort throughout five epidemic waves of SARS-CoV-2 in Rio de Janeiro, Brazil.

Since May 2020, we have been conducting a comprehensive study to understand the natural history of SARS-CoV-2 infection in Rio de Janeiro, Brazil. Our focus has been on following families, systematically collecting respiratory tract swabs and blood samples, monitoring symptoms, and gathering data on vaccine status. This paper aims to describe the household cohort across five epidemic waves of SARS-CoV-2, providing an overview of the collected data and a description of the epidemiological, clinical, and immunological characteristics and incidence of SARS-CoV-2 infection. Our cohort includes 691 participants from 189 households. During the five epidemic waves, we detected 606 infections. The incidence density of SARS-CoV-2 infection ranged from 4 (Delta) to 56 (B.1.1.33) per 1,000 person-week, with a peak in wave B.1.1.33 in all age groups. The seroprevalence of SARS-CoV-2 antibodies (IgG anti spike protein) varied from 37%, in the pre-VoC period, to 99%, in the Omicron period, progressively increasing after each wave in a similar manner regardless of age. As we have monitored the cohort continuously since the beginning of the pandemic, we were able to collect data across different scenarios according to the predominant lineage in circulation. Via active monitoring of families, we were able to carry out an epidemiological surveillance on SARS-CoV-2, including its variants, persistence of symptoms, and changes in immunity over time in the population, contributing to knowledge of the natural history of SARS-CoV-2 infection.

Frequency of Cysts by Immunofluorescence Assay in Human Brain Tissue of Suicide, Traffic Accident and Homicide Decedents

BACKGROUND: Several studies link toxoplasmosis with neuropsychiatric disorders. There are no previous reports of the frequency of the presence of parasite tissue cysts in autopsy cases from violent death or how is the distribution of frequency according to brain regions related with behavior (amygdala or hippocampus). The immune fluorescent antibody technique (IFAT) detects specific target antigens that allow the observation of bradyzoites within tissue cysts. We describe the frequency of tissue cysts using the IFAT detection in human brain tissue samples from immunocompetent persons deceased by suicide, traffic accidents and homicide. METHODS: 21 brains from the National Institute of Legal Medicine and Forensic Sciences chapter “Eje Cafetero” (including cities of Armenia, Pereira, and Manizales) of violently deceased (which includes suicide, traffic accidents and homicide) persons were obtained, subsequently sections of the amygdala and hippocampus areas were processed in paraffin and thick sections were cut from paraffin blocks and mounted on slides by means of the IFAT, by using specific monoclonal anti BAG1 protein of the bradyzoite. Serum samples from the decedents were also examined for the presence of IgG and IgM antibodies to establish the existence of a previous infection. Data such as age, gender, sociodemographic data, and type of death were also obtained. Evaluation of the differences in the percentage of positivity according to the cause of death were assayed by using the Fisher exact test. RESULTS: Two decedents (9.5%) were female and 19 (90.4%) were male. The age ranged from 14 to 65 years; most completed or were in high school level of education. By city, 9 were from Armenia (42.8%%) and 12 from Manizales (57.1%). By type of death, 11 samples (52.3%) were victims of homicide, 9 samples (42.8%) were suicide, and one sample (4.7%) was a traffic accident. We identified tissue cysts containing the bradyzoites in two cases in amygdala (9.5%), in amygdala and hippocampus in tree (14.2%), and one case only in hippocampus (4.7%). In total we have six of 23 (26%) were positive for cysts in the brain. No statistically significant differences were found between groups in the percent of positivity. After counting we found a mean 1,3 ± 0,4 cysts by 20 microscopy fields of positive brain tissue examined. We found one of 21 positives for IgG antibodies in blood (47%). Within six cases with positive by IFAT in brain tissue five were also positive in antibodies (83%) only one case was negative for antibodies. CONCLUSION: The presence of bradyzoite was demonstrated in brain tissue samples from immunocompetent patients who suffered a violent death in the cities of Armenia and Manizales, Colombia. The results suggest that the bradyzoite after infection is permanently localized in brain tissue. A higher prevalence of bradyzoite parasites is observed in amygdala samples, an important region for behavior control.

SARS-CoV-2 Post Vaccinated Adverse Effects and Efficacy in the Egyptian Population.

Vaccines are the solution to overcome SARS-CoV-2. This study aimed to determine the post-Sinopharm vaccine safety-profile and immunity through antibody titers. Data were collected using a structured questionnaire from Egyptian participants who received two doses of Sinopharm vaccine. Data were divided into three parts, the first and second parts were to detect participants’ post-first and second dose symptoms and practices, and the third for the results of IgG anti spike protein antibodies test and laboratory tests. Pain, redness, swelling at the injection site, headache, fatigue, and lethargy were the most common post-vaccine symptoms for both first and second doses. Most of the participants felt mild or no symptoms after vaccination. The symptoms started mostly during the first day post-vaccination and lasted for no more than two days. Forty-nine percent of the participants resulted in positive antibodies tests on day 18 post-vaccination. The average antibody level for vaccinated participants with past SARS-CoV-2 infection was much higher than that for non-past infected participants. These vaccines’ administration methods need to be reevaluated by changing the dose, dose interval, adding a third dose, or mixing it with other vaccines with different techniques to improve their protection rates. Further studies are required to validate this finding.

A Novel Bispecific Antibody Targeting PD-L1 and VEGF With Combined Anti-Tumor Activities.

Therapeutic monoclonal antibodies (mAbs) blocking immune checkpoints have been mainly used as monotherapy. Recently, combination therapy targeting multiple immune checkpoints has recently been explored to increase anti-cancer efficacy. Particularly, a single molecule targeting more than one checkpoints has been investigated. As dual blocking of PD-1/PD-L1 and VEGF/VEGFR has demonstrated synergism in anti-tumor activities, we developed a novel bispecific antibody, termed HB0025, which is formed via fusing the domain 2 of vascular endothelial growth factor receptor 1 (VEGFR1D2) and anti-PD-L1 mAb by using mAb-Trap technology. HB0025 almost completely retains the binding affinities and the biological activities in-vitro when compared with the parent anti-PD-L1 mAb or VEGFR1D2 fusion protein. Preclinical data demonstrated that HB0025 was more effective in inhibiting cancer growth than anti PD-L1 mAb or VEGFR1D2 fusion protein. Thus, our bispecific antibody may bring about greater clinical benefits and broader indications.

A robust mussel-inspired zwitterionic coating on biodegradable poly(L-lactide) stent with enhanced anticoagulant, anti-inflammatory, and anti-hyperplasia properties

Endowing cardiovascular stents with multiple functionalities, such as anti–proliferative, and anti–inflammatory properties, is important for combating restenosis after stent implantation. The surface properties of stents play a key role in obtaining the desired post–implantation performance. Zwitterionic coatings are resistant to non–specific adsorption, and here, a mussel–inspired zwitterionic poly (carboxybetaine acrylate–co–dopamine methacrylate) copolymer (PCBDA) was synthesized and applied to the surface of a biodegradable poly(l–lactide) stent. To obtain a zwitterionic polymer coating, the surface of a PLA stent was pretreated by co–depositing polydopamine (PDA) and polyethyleneimine (PEI). The mussel–inspired catechol moieties in the PCBDA copolymer cross–linked with the PDA–PEI hybrid to form a robust PCBDA/PDA–PEI coating on the PLA stent. The zwitterionic PCBDA formed a hydration layer once it contacted blood, preventing the non–specific adsorption of serum proteins, which further blocked subsequent coagulation and inflammation. The anti–protein adsorption tests showed that the PCBDA/PDA–PEI coating resisted the non–specific adsorption of fibrinogen, which was a positive signal for enhanced anti–coagulation. Moreover, the platelet adhesion/activation tests confirmed its antithrombotic properties. The in vivo tissue response was also greatly suppressed due to the inhibited recognition of PCBDA/PDA–PEI–coated implants by macrophages. Interestingly, unlike traditional zwitterionic polymers, due to the introduction of adhesive catechol moieties, PCBDA supported the growth of endothelial cells, demonstrating its ability to selectively direct the fate of endothelial cells and smooth muscle cells. In vivo stent implantation results showed inhibited intimal hyperplasia due to the protection of PCBDA/PDA–PEI. Overall, due to the specific characteristics of the mussel–inspired PCBDA/PDA–PEI coating, anticoagulation, anti–inflammatory, and anti–proliferation properties were obtained; thus, it is expected that this coating can be used to inhibit restenosis and realize endothelialization after stent implantation.

Impact of DOTA Conjugation on Pharmacokinetics and Immunoreactivity of [177Lu]Lu-1C1m-Fc, an Anti TEM-1 Fusion Protein Antibody in a TEM-1 Positive Tumor Mouse Model

1C1m-Fc, an anti-tumor endothelial marker 1 (TEM-1) scFv-Fc fusion protein antibody, was previously successfully radiolabeled with 177Lu. TEM-1 specific tumor uptake was observed together with a non-saturation dependent liver uptake that could be related to the number of dodecane tetraacetic acid (DOTA) chelator per 1C1m-Fc. The objective of this study was to verify this hypothesis and to find the best DOTA per 1C1m-Fc ratio for theranostic applications. 1C1m-Fc was conjugated with six concentrations of DOTA. High-pressure liquid chromatography, mass spectrometry, immunoreactivity assessment, and biodistribution studies in mice bearing TEM-1 positive tumors were performed. A multi-compartment pharmacokinetic model was used to fit the data and a global pharmacokinetic model was developed to illustrate the effect of liver capture and immunoreactivity loss. Organ absorbed doses in mice were calculated from biodistribution results. A loss of immunoreactivity was observed with the highest DOTA per 1C1m-Fc ratio. Except for the spleen and bone, an increase of DOTA per 1C1m-Fc ratio resulted in an increase of liver uptake and absorbed dose and a decrease of uptake in tumor and other tissues. Pharmacokinetic models correlated these results. The number of DOTA per antibody played a determining role in tumor targeting. One DOTA per 1C1m-Fc gave the best pharmacokinetic behavior for a future translation of [177Lu]Lu-1C1m-Fc in patients.

PREPARATION, OXYGEN PERMEABILITY, OPTICAL TRANSPARENCY,WATER CONTENT,PROTEIN ADSORPTION AND EVALUATION OF BACTERIAL ADHESION PROPERTIES OF NEW SILICONE HYDROGEL THROUGH MODULATING SILICONE AND HYDROPHILIC MONOMER

Silicone Hydrogel (SiHy) contact lenses are extremely fruitful in compare is onto preceding soft lenses; they were established to offer superior oxygen permeability. In the following paper, the hydrogel contact lenses based on silicone were made through the polymerization of 3- (methacryloyloxy) propyltris(trimethylsiloxy) silane (TRIS) with N,N dimethylacryl amide (DMA), Methyl methacrylate (MMA),and 2-hydroxyethyl methacrylate (HEMA). The features of silicone hydrogel lenses were studied depending on the approaches like equilibrium water component, oxygen permeability, optical transparency, protein adsorption, as well as cell toxicity.The outcomes revealed which the TRIS component in all makings rose the oxygen penetrability and decreased the equilibrium water component, while the DMA& HEMA participated the hydrophilicity of the silicon hydrogels and increases the equilibrium water content (EWC)%. The higher worth of oxygen penetrability was 102barrers, conforming to an equilibrium made of water component of 66%. generally, the silicone hydrogels in this work displayed decent oxygen permeability as well as optical transparency and anti protein adsorption. Hence forth, those polymers made of silicone hydrogel might be possible for manufacturing contact lens.

Integrated strategy for the separation of endotoxins from biofluids. LPS capture on newly synthesized protein

Endotoxin, or lipopolysaccharide (LPS), is recognized as the most powerful microbial mediator implicated in the pathogenesis of sepsis, a life-threatening organ disfunction whose overall prognosis is still poor. Therefore, the need to detect and remove LPS has led some authors to focus their efforts on the design of pathogen cleansing microdevices. Micromagnetophoretic cleansing involves a first step for the selective binding and sequestration of LPS on non-toxic particles followed by the removal of the loaded particles from the fluid medium. While outstanding progress has been made in magnetic separation, the state of the art of the first step still needs to fill important gaps in knowledge to advance towards the development of detection systems. Thus, this work reports an integrated methodology to advance the design of the LPS sequestration stage to promote its separation from biofluids. The methodology combines protein and separation fundamentals and starts with the synthesis of an anti LPS factor protein (LALF), followed by the quantitative determination of its binding strength to LPS. For this analysis LALF was supported on agarose beads and the variables affecting LPS binding, as binding and capture temperature, the optimum bead:protein and protein:LPS ratios, have been experimentally studied. The methodology and results here reported constitute the information needed to advance the knowledge for the correct design of LPS separation devices.

Enhanced anti–protein fouling of PVDF membrane via hydrophobic–hydrophobic adsorption of styrene–terminated amphiphilic linker

The amphiphilic copolymer polystyrene-maleic anhydride (SMA)-2-hydroxyethyl acrylate (HEA) was synthesized for the first time in this study, and used as a linker to graft zwitterionic sulfobetaine methacrylate (SBMA) in order to enhance the hydrophilicity and anti-adsorption ability of polyvinylidene difluoride (PVDF) membranes. This amphiphile linker is composed of a hydrophobic extremity, i.e., the SMA copolymer, and a hydrophilic extremity, i.e., the HEA. The SMA-HEA linker was first dispersed into the PVDF matrix via the blending method, and the hydrophobic polystyrene of SMA was adsorbed into the PVDF membrane via hydrophobic–hydrophobic interaction. The SBMA was then grafted onto the expanded HEA via free radical polymerization. The functional groups, morphologies, and hydrophilicity of the amphiphile-modified PVDF membranes were analyzed by attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR), field emission scanning electron microscopy (FE-SEM), and water contact angle (WCA) measurements. The PVDF membrane modified with the SMA-HEA amphiphile linker showed significant enhancement in hydrophilicity as the WCA decreased from 135.0° to 45.0°, and the protein adsorption of the modified membrane decreased from 112.5 to 33.3μg⋅cm−2. Furthermore, dynamic protein separation was performed, and the pure water flux recovery rate reached 100%, indicating good anti-adsorption ability.

Alcoholic Extract of Shilajit as Anti Protein Denaturation, Anti Blood Hemolysis, and Anti Microbial

Alcoholic Extract of Shilajit as Anti Protein Denaturation, Anti Blood Hemolysis, and Anti Microbial

How fermentation by lactic acid bacteria can address safety issues in legumes food products?

Fermented Asian foods have recently gained popularity, crossing from Asian communities to mainstream markets, in many western countries. However, less attention has been paid to the safety of these foods. In South-East Asia, fermented products are still produced following traditional methods. Therefore, consumers are not confident in their microbial safety. The challenges awaiting fermentation in South-East Asia are thus to improve safety and quality in a sustainable system producing tasty and typical fermented products. A possible solution could be the use of starter cultures able to increase the safety of food stuffs. Starters could also help to standardize the production process and reduce ripening times. The current review addresses the role of lactic acid bacteria on the microbiological and chemical safety of Asian legume based fermented products. In particular, their role in the reduction of anti nutritional compounds (e.g. phytates) and protein allergenicity is discussed. Moreover, starters can inihibit the development of amino acid-decarboxylating microbes preventing the accumulation of biogenic amines, they can also be useful to reduce the accumulation of mycotoxins and inhibit pathogens’ development. Finally, their role in the degradation of pesticides is analyzed.

Targeting Alpha Synuclein Aggregates in Cutaneous Peripheral Nerve Fibers by Free-floating Immunofluorescence Assay.

To date, for most neurodegenerative diseases only a post-mortem histopathological definitive diagnosis is available. For Parkinson's disease (PD), the diagnosis still relies only on clinical signs of motor involvement that appear later on in the disease course, when most of the dopaminergic neurons are already lost. Hence, there is a strong need for a biomarker that can identify patients at the beginning of disease or at the risk of developing it. Over the last few years, skin biopsy has proved to be an excellent research and diagnostic tool for peripheral nerve diseases such as small fiber neuropathy. Interestingly, a small fiber neuropathy and alpha synuclein (αSyn) neural deposits have been shown by skin biopsy in PD patients. Indeed, skin biopsy has the great advantage of being an easily accessible, minimally invasive and painless procedure that allows the analysis of peripheral nervous tissue prone to the pathology. Moreover, the possibility of repeating the skin biopsy in the course of the follow-up of the same patient allows studying the longitudinal correlation with the disease progression. We set up a standardized reliable protocol to investigate the presence of αSyn aggregates in skin nerve fibers of the PD patient. This protocol involves few short fixation steps, a cryotome sectioning and then a free-floating immunofluorescence double-staining with two specific antibodies: anti Protein Gene Product 9.5 (PGP9.5) to mark the cutaneous nerve fibers and anti 5G4 for detecting αSyn aggregates. It is a versatile, sensitive and easy to perform protocol that can also be applied for targeting other proteins of interest in skin nerves. The ability to mark αSyn aggregates is another step forward to the use of skin biopsy as a tool for establishing a pre-mortem histopathological diagnosis of PD.

Detection of Brucella abortus by immunofluorescence assay using anti outer membrane protein of 19 kDa antibody.

Brucellosis in humans is one of the most prevalent zoonotic diseases around the world with more than 500,000 new cases per year. It is a weakening disease that requires long-term antibiotic treatment, often resulting in permanent and disabling consequences. Outer membrane proteins (OMPs) of Brucella, which are non-lipopolysaccharide (LPS) antigens, have been used for the diagnostic kits of brucellosis and vaccine design. The aim of this study was to identify Brucella abortus with an immunofluorescence (IF) test using an antibody against recombinant outer membrane protein (OMP) of 19 kDa of this bacterium. The OMP19 gene of Brucella spp. was synthesized, cloned and expressed in Escherichia coli cells. The OMP19 protein was purified by metal chelate affinity chromatography and subsequently used for the immunization of rabbits to produce a polyclonal antibody. Then, this antibody was conjugated to fluorescein isothiocyanate (FITC) and used for the detection of Brucella by an IF test. Also, the sensitivity and specificity of this antibody for the diagnosis of clinical isolates was calculated. Outer membrane protein 19 was expressed well and reacted with a commercial antiserum against His-tag in an immunoblot assay. Polyclonal antibodies obtained from the serum of rabbits immunized with the purified protein showed strong reactivity in the enzyme-linked immunosorbent assay (ELISA). Moreover, the polyclonal antibody conjugated to FITC was able to properly identify Brucella abortus. Sensitivity and specificity of this IF test in comparison with a polymerase chain reaction (PCR) assay was 84.2% and 50%, respectively. This high-titer antibody could potentially be valuable for the specific diagnostic test of brucellosis.

Studies on the preparation and controlled release of redox/pH-responsive zwitterionic nanoparticles based on poly-L-glutamic acid and cystamine

The enhancement of tumor intracellular drug uptake and resistance against nonspecific protein adsorption are essential for an injectable anticancer drug carrier. In the present study, a new type of redox/pH-responsive zwitterionic nanoparticles (NPs) was prepared using poly-L-glutamic acid and cystamine in aqueous solutions under mild conditions. The NPs showed surface charge convertible feature in response to pH change of the solutions. The NPs demonstrated excellent anti nonspecific protein adsorption. In vitro release profiles of the NPs, they showed redox/pH dual sensitivities in vitro release. The effective intracellular delivery behaviors were verified through investigation of cell viability, and confocal laser scanning microscopy observation of HeLa cells after incubation with the DOX-loaded NPs. The NPs were non-cytotoxic and would have potential applications as a drug delivery vehicle for enhancing intracellular uptake of anticancer drugs.

Anti-ROR1 scFv-EndoG as a Novel Anti-Cancer Therapeutic Drug

Aim:Immunotoxins are proteins that consist of an antibody fragment linked to a toxin, used as agents for targeted therapy of cancers. Although the most potent immunotoxins are made from bacterial and plant toxins, obstacles which contribute to poor responses are immunogenicity in patients and rapid development of neutralizing antibodies. In the present study we proposed a new therapeutic immunotoxin for targeted cancer therapy of ROR1 expressing cancers: an anti ROR1 single chain fragment variable antibody (scFv)-endonuclease G (anti ROR1 scFv-EndoG).Methods:The three-dimensional structure of anti ROR1 scFv-EndoG protein was modeled and structure validation tools were employed to confirm the accuracy and reliability of the developed model. In addition, stability and integrity of the model were assessed by molecular dynamic (MD) simulation.Results:All results suggested the protein model to be acceptable and of good quality.Conclusions:Anti-ROR1 scFv-EndoG would be expected to bind to the ROR1 extracellular domain by its scFv portion and selectively deliver non-immunogenic human endonuclease G enzyme as an end-stage apoptosis molecule into ROR1-expressing cancer cells and lead rapidly to apoptosis. We believe that anti ROR1 and other anti-tumor antigen scFv-EndoG forms may be helpful for cancer therapy.

The Study of Antioxidant, Membrane Stabilization, Anti Protein-Denaturation Property and Analysis of Phytochemicals in Three Species of Tagetes Leaf and Flower Extract

Since the ancient time nature provides several herbal phyto-chemicals for the beneficial of human. Tagetes sp is one of the common and valuable herbs with multiple uses. In the current study, qualitative screening of different phytoconstituents, anti protein denaturation and membrane stabilization property was checked in ethanolic extract of three different species of Tagetes sp namely Tagetes petula, Tagetes tenuifolia, Tagetes erecta. Three different concentration 50, 75, 100 microlitre were taken for membrane stabilization and anti protein denaturation method using 50 microgram/ml sodium diclofenec as a reference drug. The study revealed that flower and leaf extract from each sample have several bioactive compounds and significant membrane stabilization and anti protein denaturation property. Presence of several phyto-chemicals may help to scavenge the reactive oxygen, potent membrane destabilizing and anti protein denaturation agent in human. So, in future it could be possible to develop a new phyto derived drug for an inflammatory disease in human as literature review shows that the membrane destabilization and protein denaturation are the principal cause of inflammation related disease.

Advances in management of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the leading causes of death by cancer worldwide due to a dismal prognosis. The aim of this review is to summarize the main advances in the pathophysiology and management of HCC. Genomic analysis has recently delineated the key signaling pathways aberrantly deregulated in HCC (telomere maintenance, cell cycle gene, Wnt/β-catenin, epigenetic modifier, oxidative stress etc.). Major advances in the clinical care of patients with HCC are helping to refine the diagnosis algorithm and tumor staging. Extension of criteria for liver transplantation, but also for liver resection and percutaneous ablation, aims to increase the number of patients being treated in a curative attempt. Moreover, radioembolization is a competitor for transarterial chemoembolization in Barcelona clinic liver cancer B patients, and sorafenib in Barcelona clinic liver cancer C with tumor portal thrombosis. In advanced HCC, sorafenib is the standard of first-line care and regorafenib as a second line. New concepts on liver resection, percutaneous ablation or liver transplantation aim to extend the numbers of patients treated for HCC in curative attempts. Moreover, immunotherapy (anti death protein 1) and biotherapy adapted to tumor biology seem promising and are currently being tested in advanced stages.

Development of biohybrid immuno-selective membranes for target antigen recognition

Membranes are gaining increasing interest in solid-phase analytical assay and biosensors applications, in particular as functional surface for bioreceptors immobilization and stabilization as well as for the concentration of target molecules in microsystems.In this work, regenerated cellulose immuno-affinity membranes were developed and they were used for the selective capture of interleukin-6 (IL-6) as targeted antigen. Protein G was covalently linked on the membrane surface and it was successfully used for the oriented site-specific antibody immobilization. The antibody binding capacity of the protein G-coupled membrane was evaluated. The specific anti IL-6 antibody was immobilized and a quantitative analysis of the amount of IL-6 captured by the immuno-affinity membrane was performed. The immobilization procedure was optimized to eliminate the non-specific binding of antigen on the membrane surface. Additionally, the interaction between anti IL-6 antibody and protein G was stabilized by chemical cross-linking with glutaraldehyde and the capture ability of immuno-affinity membranes, with and without the cross-linker, was compared. The maximum binding capacity of the protein G-coupled membrane was 43.8µg/cm2 and the binding efficiency was 88%. The immuno-affinity membranes showed a high IL-6 capture efficiency at very low antigen concentration, up to a maximum of 91%, the amount of captured IL-6 increased linearly as increasing the initial concentration. The cross-linked surface retained the antigen binding capacity demonstrating its robustness in being reused, without antibody leakage or reduction in antibody binding capacity. The overall results demonstrated the possibility of a reliable application of the immuno-affinity membrane developed for biosensors and bioassays also in multiple use.

Evaluation of diagnostic role of IP-10 (Interferon Gamma Inducible Protein) and anti PGL-1 antibody in pediatric leprosy

Background: Leprosy is among the world’s oldest and most dreaded diseases and it has been synonymous with stigma and discrimination due to the hideous deformities it produced, mystery around its aetiology and transmission and lack of any effective remedy till recently. Leprosy is characterized by a long and variable incubation period and a chronic clinical course. Diagnosis of leprosy is essentially based on clinical features. Although the majority of cases can be diagnosed clinically yet alternative methods for diagnosis are required especially for early cases. The present study is aimed to assess the diagnostic value of anti PGL-1 antibody and Interferon Gamma Inducible Protein (IP-10) and to compare these techniques with skin smear. Materials and Methods: A prospective study was done on 30 patients below 16 years of age between March 2014 to March 2015. Results: In this study 13 / 30 (43.33%) cases were positive by Anti PGL-1 antibody based ELISA and 12/30 (40%) by IP-10 based ELISA against 9/ 30 (30.00%) cases by skin smears for AFB. Hence Anti PGL-1 and IP-10 based ELISA has more diagnostic value than slit skin smear for AFB in confirmation of leprosy cases. Conclusion: This study supports that Anti PGL-1 antibody and Interferon Gamma induced protein (IP-10) enhance the diagnostic yield of leprosy when compared to routine skin smears stained by Z.N staining. They are important diagnostic tools for definitive diagnosis in early cases of leprosy.