Purification and immunogenic characterization of anti-LPS Rabbit antibodies produced against Vibrio cholerae inaba O1 extracted Lipopolysaccharide

Abstract

Conclusion: This research showed that controlling postprandialhyperglycemia with appropriate dose of insulin injection and foodrestriction in diabetic rats causes reduced serum triglyceride. It may besuggested that prevention of postprandial hyperglycemia and foodrestriction may decrease vascular complications due to hypertriglycer-idemia in diabetes.Keywords: Postprandial hyperglycemia, Diabetes, Cardiovasculardisorder, Triglyceride, Food restriction (FR)doi:10.1016/j.clinbiochem.2011.08.332Poster – [A-10-274-2]Purification and immunogenic characterization of anti-LPS Rabbitantibodies produced against Vibrio cholerae inaba O1 extractedLipopolysaccharideWalead Ebrahimi zadeh, Seyedlatif Mousavi gargari,Masoumeh Rajabi bazl, Mohammad MohammadiShahed University, Qom, IranE-mail addresses: v_ebrahimi@hotmail.com (W. Ebrahimi zadeh),slmousavi@shahed.ac.ir (S. Mousavi gargari),masoumeh_rajabibazl@yahoo.com (M. Rajabi bazl),mohamadi74@yahoo.com (M. Mohammadi)Introduction: Vibrio cholerae can cause cholera, which is an acute,diarrheal illness that can result in severe dehydration and even deathwithin a matter of hours. The bacterium is mainly subdivided by its Oantigen ( Lipopolysaccharide). LPS is one of the key virulence factorsof these bacteria which makes it a good candidate for production ofantibody vaccines. In this study the V. cholerae was neutralized withIgG purified from immunized rabbit with LPS, and then used tochallenge BALB/c mice.Materials and methods: V. cholerae inaba O1 lipopolysaccharidewas extracted via hot phenol/water method and assayed withTricine-SDS-PAGE and LPS-Silver stain methods. After immunizationof rabbits with LPS, blood was collected and assayed for antibodyproduction by ELISA. Antibodies were first precipitated by ammo-nium sulfate salt-out and then purified by DEAE cellulose chromatog-raphy. Affinity potential of antibodies to LPS and V. cholerae wasstudied by ELISA. V. cholerae was incubated with antibodies at 37°Cfor 1 h and gavaged into BALB/c mice stomach. Control mice wereonly given same amount of bacterium.Results: Extracted LPS showed similar pattern compared topositive control in Tricine-SDS-PAGE. ELISA results indicated thatantibodies have similar affinity to extracted LPS and V.cholerae.Compared to control mice, after treating the bacteria with antibodythe CFU for LD50 was significantly increased.Conclusion: Produced antibodies showed high attachment abilitynotonlytotheextractedLPSbutalsototheV.cholerae inabaO1,andwereabletomaintainthisfunctionininvivoexperimentwhichindicatesthatanti-LPS antibodies are good candidates for antibody vaccines.Keywords: Vibrio cholerae inaba O1, LPS, Antibody, ELISA, LD50, BALB/c micedoi:10.1016/j.clinbiochem.2011.08.333Poster – [A-10-280-1]Comparison of IgY extraction methodsZiba Vaise Malekshahi, Latif Mousavi Gargari Seyed,Hassan Shirazi MohamadTehran, IranE-mail addresses: Z.malekshahi@yahoo.com (Z. Vaise Malekshahi),smousavi@shahed.ac.ir (L. Mousavi Gargari Seyed),mhshirazi@tums.ac.ir (H. Shirazi Mohamad)Introduction: The production of antibodies (Abs) in chickens andthe extraction of specific Abs from egg yolk (IgYAbs) are increasinglyattracting the interest of the scientific community. Egg yolk is animportant source of antibodies.Materials and methods: After injection of UreC recombinantprotein of Helicobacter pylori bacterium to white leghorn hens,immunization of hens was confirmed by indirect ELISA technique.Produced IgY was precipitated from egg yolk, using poly ethyleneglycol,sodium sulphate and isopropanol-aceton methods. ProducedIgY assayed using indirect ELISA technique and SDS-PAGE analysis.Results: The biggest obstacle for isolation of chicken antibodies(IgY) is the removal of lipids, so better method for IgY purification ispoly ethylene glycol. The purity of our purified IgY with poly ethyleneglycol method was 70% and a yield of 9.4 mg of IgYper ml of egg yolk.Conclusion: The choice of a suitable IgY purification method isimportant. In recent years, poly ethylene glycol (PEG) precipitationmethods has become the most commonly used and most effectiveprocedure.Keywords: Antibodies, Hens, IgY extraction, PEGdoi:10.1016/j.clinbiochem.2011.08.334Poster – [A-10-298-1]Evaluating cardiovascular risk factors among type 2 diabetic andhealthy populaceHossein Pourghadamyari, Reza Parizadeh Seyed Mohammad,Shima Tavallaie, Amirhossein Sahebkar, Roghayeh Paydar,Majid Ghayour Mobarhan, Reza karamiDepartment of Phisiology, School of Medicine, Mashhad University ofMedical science, Mashhad, IranE-mail addresses: GhadamyariH871@mums.ac.ir (H. Pourghadamyari),ParizadehMR@mums.ac.ir (R. Parizadeh Seyed Mohammad),TavalaieSH1@mums.ac.ir (S. Tavallaie),SahebkarAH811@mums.ac.ir (A. Sahebkar),PaydarR1@mums.ac.ir (R. Paydar),GhayourM@mums.ac.ir (M. Ghayour Mobarhan),karamir851@mums.ac.ir (R. karami)Introduction: Type 2 diabetes mellitus is characterized byelevated blood glucose levels and increased long-term risk formorbidity and mortality, most often due to cardiovascular events.Some studies demonstrate predisposing effect of diabetes on heartattack and cardiovascular risk factors. The aim of this study iscomparing cardiovascular risk factors between diabetic and healthypeople.Methods: We studied 110 patients with type 2 diabetes and 345age and sex-matched healthy controls. The most common cardiovas-cular risk factors including fasting blood sugar (FBS), systolic bloodpressure (SBP), diastolic blood pressure (DBP) and lipid profile weremeasured. Lipid profile was estimated through measuring triglycer-ides, low-density lipoprotein cholesterol (LDL-C) and high-densitylipoprotein cholesterol (HDL-C) levels. Blood glucose level wasmeasured by autoanalayzer device and photometry method. Lipidprofile measurement was performed by some routine enzymaticmethods. The data were analyzed by Mann–Whitney test amongdiabetic and healthy people. P-value less than 0.05 was consideredsignificant.Results: Our findings demonstrate there is significant differencebetween triglyceride levels in diabetic and healthy people (p<0.05),