Small Molecular Mass Research Articles

Improved Low-field NMR porosity characterization of cementitious materials containing water-soluble organic admixtures

Water-soluble organic admixtures (WSOAs) are widely used in cementitious materials, and may affect the porosity, but their effects on low-field nuclear magnetic resonance (LF-NMR) measurement are often overlooked. Using viscosity modifying admixture (VMA) as an example, this study demonstrates the presence of WSOAs would lead to an overestimation of porosity and a leftward shift of pore size distribution using LF-NMR. This is attributed to the formation of hydrogen bonds between polymer and water molecules, and the decreased distance between nuclear spins, respectively. A strategy is proposed to improve the accuracy of the porosity characterization by eliminating the additional effects of WSOAs in LF-NMR testing. The results show that the addition of small molecular mass non-ionic VMAs replacing mixing water (within a 20 % replacing rate) does not alter the porosity of the paste. This method can be extended to characterize the porosity of various porous materials containing WSOAs.

Cancer Cell Small Molecule Secretome Induces the Immune Checkpoint NKG2A and Dysfunction of Human CD8+ T Cells.

PD-1 blockade has been approved for head and neck squamous cell carcinoma (HNSCC) patients. However, many HNSCC patients do not respond to this treatment, and other tumor microenvironmental factors may promote resistance to PD-1 blockade. We previously identified increased expression of the inhibitory receptor NKG2A on CD8+ T cells in HNSCC tumors compared with T cells in matching PBMC samples. Mechanisms that promote NKG2A expression and the role of NKG2A on human T cells in the tumor microenvironment, however, are uncertain. In this study, we show that tumor-conditioned media (TCM) of HNSCC cancer cell lines or ascites fluid from colorectal carcinoma patients is sufficient to induce the expression of NKG2A and other inhibitory receptors on activated CD8+ T cells isolated from PBMCs of healthy donors. Boiling or small molecular mass cutoff filtering did not eliminate the effect of TCM, suggesting that a small molecule promotes NKG2A. T cell activation in TCM decreased the basal and maximal mitochondrial respiration to metabolically restrain CD8+ T cells. Functionally, T cell activation in TCM reduced CD8+ T cell cytotoxicity as shown by lower production of cytokines, granzyme B, and perforin. Furthermore, TCM prevented CD8+ T cells from killing cancer cells in response to an anti-CD19/anti-CD3 bispecific T cell engager. Thus, a small secreted molecule from HNSCC cells can induce NKG2A expression and promote T cell dysfunction. Our findings may lead to targets for novel cancer therapies or biomarkers for NKG2A blockade response and provide a model to study T cell dysfunction and impaired metabolism.

A sensitive Raman spectroscopy sensor for determination cardiac troponin I based on proteolytic peptide magnetic imprinting technology

This paper proposes a selective strategy for quantitative detecting cardiac troponin I (cTnI) in clinical serum samples by using Raman spectroscopy combined with magnetic molecularly imprinted polymers (MMIPs). The signature peptide was selected as a template for molecular imprinting due to its chemical stability and small molecular mass. MMIPs were synthetized with the signature peptide as template and methylacrylic acid as monomer in presence of vinyl-modified magnetic particles. MMIPs with high magnetic character and adsorption properties were able to be stained by Coomassie Brilliant Blue G-250 after the target template was specifically captured, which showing a characteristic absorption peak at Raman shift 1610 cm−1. The Raman signal intensity at 1610 cm−1 showed a linear relationship in the 0.01 to 100 ng·mL−1 concentration range. The recovery of cTnI in serum were 92.7–111.2 % with relative standard deviation in the range of 2.51 % to 8.52 %. This paper has important practical application for preliminary rapid detection of cTnI in clinical serum samples.

Detection of Mycobacterium tuberculosis complex field infections in cattle using fecal volatile organic compound analysis through gas chromatography-ion mobility spectrometry combined with chemometrics.

Bovine tuberculosis is considered a re-emerging disease caused by different species from the Mycobacterium tuberculosis complex (MTC), important not only for the livestock sector but also for public health due to its zoonotic character. Despite the numerous efforts that have been carried out to improve the performance of the current antemortem diagnostic procedures, nowadays, they still pose several drawbacks, such as moderate to low sensitivity, highlighting the necessity to develop alternative and innovative tools to complement control and surveillance frameworks. Volatilome analysis is considered an innovative approach which has been widely employed in animal science, including animal health field and diagnosis, due to the useful and interesting information provided by volatile metabolites. Therefore, this study assesses the potential of gas chromatography coupled to ion mobility spectrometry (GC-IMS) to discriminate cattle naturally infected (field infections) by MTC from non-infected animals. Volatile organic compounds (VOCs) produced from feces were analyzed, employing the subsequent information through chemometrics. After the evaluation of variable importance for the projection of compounds, the final discriminant models achieved a robust performance in cross-validation, as well as high percentages of correct classification (>90%) and optimal data of sensitivity (91.66%) and specificity (99.99%) in external validation. The tentative identification of some VOCs revealed some coincidences with previous studies, although potential new compounds associated with the discrimination of infected and non-infected subjects were also addressed. These results provide strong evidence that a volatilome analysis of feces through GC-IMS coupled to chemometrics could become a valuable methodology to discriminate the infection by MTC in cattle. IMPORTANCE Bovine tuberculosis is endemic in many countries worldwide and poses important concerns for public health because of their zoonotic condition. However, current diagnostic techniques present several hurdles, such as low sensitivity and complexity, among others. In this regard, the development of new approaches to improve the diagnosis and control of this disease is considered crucial. Volatile organic compounds are small molecular mass metabolites which compose volatilome, whose analysis has been widely employed with success in different areas of animal science including animal health. The present study seeks to evaluate the combination of fecal volatilome analysis with chemometrics to detect field infections by bovine tuberculosis (Mycobacterium tuberculosis complex) in cattle. The good robust performance of discriminant models as well as the optimal data of sensitivity and specificity achieved highlight volatilome analysis as an innovative approach with huge potential.

First Results from the JWST Early Release Science Program Q3D: The Warm Ionized Gas Outflow in z ∼ 1.6 Quasar XID 2028 and Its Impact on the Host Galaxy

Quasar feedback may regulate the growth of supermassive black holes, quench coeval star formation, and impact galaxy morphology and the circumgalactic medium. However, direct evidence for quasar feedback in action at the epoch of peak black hole accretion at z ≈ 2 remains elusive. A good case in point is the z = 1.6 quasar WISEA J100211.29+013706.7 (XID 2028), where past analyses of the same ground-based data have come to different conclusions. Here, we revisit this object with the integral-field unit of the Near Infrared Spectrograph on board the JWST as part of Early Release Science program Q3D. The excellent angular resolution and sensitivity of the JWST data reveal new morphological and kinematic substructures in the outflowing gas plume. An analysis of the emission-line ratios indicates that photoionization by the central quasar dominates the ionization state of the gas with no obvious sign for a major contribution from hot young stars anywhere in the host galaxy. The rest-frame near-UV emission aligned along the wide-angle cone of outflowing gas is interpreted as a scattering cone. The outflow has cleared a channel in the dusty host galaxy, through which some of the quasar ionizing radiation is able to escape and heat the surrounding interstellar and circumgalactic media. Although the warm ionized outflow is not powerful enough to impact the host galaxy via mechanical feedback, radiative feedback by the active galactic nucleus, aided by the outflow, may help to explain the unusually small molecular gas mass fraction in the galaxy host.

Dissymmetric Triaryltriazines: Small Mass Columnar Glasses.

Columnar liquid crystals with very small molecular masses that form anisotropic glasses well above room temperature are obtained by mixed dissymmetric substitution of sym-triazine with ester-bearing phenyl and phenanthryl or tetrahelicenyl moieties. The combination of low molecular symmetry with configurational flexibility and short polar ester moieties stabilizes the mesophase over large temperature ranges and induces pronounced calorimetric glass transitions within the anisotropic fluid despite the smallness of the molecules. In contrast to more symmetrical homologs, no ester tails longer than ethyl are necessary to induce the liquid crystalline state, allowing for the near-absence of any insulating and weight-increasing alkyl periphery. Films drop-cast from solution show in all cases emission spectra that do not show significant change of fluorescence emission upon annealing, indicating that the columnar hexagonal mesoscopic order is obtained directly upon deposition from solution and is resistant to crystallization upon annealing.

Removal of tar from waste textiles by molten carbonates pyrolysis in bubbling reactor

Nowadays, the amount of waste textiles (WT) was increasing due to the increase in population, thermochemical processes owe the potential to process WT on large-scale. However, tar removal is the key to industrial thermochemical treatment. Molten carbonates pyrolysis technology is proposed to remove WT tar in this study. The result showed that molten carbonates favor catalytic reaction medium for turning tar into char and small molecular pyrolysis gases (including H2, CO, and CO2), meanwhile, excellent heat and mass transfer performance of molten carbonates can also reinforce the reaction. 94.95 wt% of tar was removed at 550 °C with flow rate of 0.15 L/min. Molten carbonates not only promoted decarboxylation of acid in tar, but also enhanced the reaction between coke and carbon dioxide, which in return benefits lifetime of molten carbonates, so after recycling eight times, tar removal rate was still above 90 %. The kinetic study suggested that lower flow rate laid positive influence on tar elimination by contributing to mass and heat transfer inside the bubbles.

Toxic or beneficial? What is the role of food‐relevant selenium species selenoneine?

Toxic or beneficial? What is the role of food‐relevant selenium species selenoneine?

Advanced Proteomic and Bioinformatic Tools for Predictive Analysis of Allergens in Novel Foods.

In recent years, novel food is becoming an emerging trend increasingly more demanding in developed countries. Food proteins from vegetables (pulses, legumes, cereals), fungi, bacteria and insects are being researched to introduce them in meat alternatives, beverages, baked products and others. One of the most complex challenges for introducing novel foods on the market is to ensure food safety. New alimentary scenarios drive the detection of novel allergens that need to be identified and quantified with the aim of appropriate labelling. Allergenic reactions are mostly caused by proteins of great abundance in foods, most frequently of small molecular mass, glycosylated, water-soluble and with high stability to proteolysis. The most relevant plant and animal food allergens, such as lipid transfer proteins, profilins, seed storage proteins, lactoglobulins, caseins, tropomyosins and parvalbumins from fruits, vegetables, nuts, milk, eggs, shellfish and fish, have been investigated. New methods for massive screening in search of potential allergens must be developed, particularly concerning protein databases and other online tools. Moreover, several bioinformatic tools based on sequence alignment, motif identification or 3-D structure predictions should be implemented as well. Finally, targeted proteomics will become a powerful technology for the quantification of these hazardous proteins. The ultimate objective is to build an effective and resilient surveillance network with this cutting-edge technology.

Lithium poisoning and renal replacement therapy: pathophysiology and current clinical recommendations.

Lithium intoxication is still an undefined and underestimated disease, especially those cases requiring extracorporeal treatment. Lithium is a monovalent cation with small molecular mass of 7Da that has been regularly and successfully used since 1950 in the treatment of mania and bipolar disorders. However, its careless assumption can lead to a wide spectrum of cardiovascular, central nervous system and kidney diseases in case of acute, acute on chronic and chronic intoxications. In fact, lithium serum range is strict between 0.6 and 1.3mmol/L, with a mild lithium toxicity observed at the steady-state of 1.5-2.5mEq/L, moderate toxicity when lithium reaches 2.5-3.5mEq/L, and severe intoxication with observed serum levels > 3.5mEq/L. Its favorable biochemical profile allows the complete filtration and partial reabsorption in the kidney due to the similarity to sodium and also the complete removal by renal replacement therapy, that should be considered in specific poisoning conditions. In this narrative and updated review we discussed a clinical case of lithium intoxication, the different pattern of diseases attributable to excessive lithium load and the current indications for extracorporeal treatment.

Mitochondria-containing extracellular vesicles (EV) reduce mouse brain infarct sizes and EV/HSP27 protect ischemic brain endothelial cultures.

Ischemic stroke causes brain endothelial cell (BEC) death and damages tight junction integrity of the blood-brain barrier (BBB). We harnessed the innate mitochondrial load of BEC-derived extracellular vesicles (EVs) and utilized mixtures of EV/exogenous 27 kDa heat shock protein (HSP27) as a one-two punch strategy to increase BEC survival (via EV mitochondria) and preserve their tight junction integrity (via HSP27 effects). We demonstrated that the medium-to-large (m/lEV) but not small EVs (sEV) transferred their mitochondrial load, that subsequently colocalized with the mitochondrial network of the recipient primary human BECs. Recipient BECs treated with m/lEVs showed increased relative ATP levels and mitochondrial function. To determine if the m/lEV-meditated increase in recipient BEC ATP levels was associated with m/lEV mitochondria, we isolated m/lEVs from donor BECs pre-treated with oligomycin A (OGM, mitochondria electron transport complex V inhibitor), referred to as OGM-m/lEVs. BECs treated with naïve m/lEVs showed a significant increase in ATP levels compared to untreated OGD cells, OGM-m/lEVs treated BECs showed a loss of ATP levels suggesting that the m/lEV-mediated increase in ATP levels is likely a function of their innate mitochondrial load. In contrast, sEV-mediated ATP increases were not affected by inhibition of mitochondrial function in the donor BECs. Intravenously administered m/lEVs showed a reduction in brain infarct sizes compared to vehicle-injected mice in a mouse middle cerebral artery occlusion model of ischemic stroke. We formulated binary mixtures of human recombinant HSP27 protein with EVs: EV/HSP27 and ternary mixtures of HSP27 and EVs with a cationic polymer, poly (ethylene glycol)-b-poly (diethyltriamine): (PEG-DET/HSP27)/EV. (PEG-DET/HSP27)/EV and EV/HSP27 mixtures decreased the paracellular permeability of small and large molecular mass fluorescent tracers in oxygen glucose-deprived primary human BECs. This one-two punch approach to increase BEC metabolic function and tight junction integrity may be a promising strategy for BBB protection and prevention of long-term neurological dysfunction post-ischemic stroke.

On the Physical Stability of Leucine-Containing Spray-Dried Powders for Respiratory Drug Delivery.

Carrier-free spray-dried dispersions for pulmonary delivery, for which the demand is growing, frequently require the incorporation of dispersibility-enhancing excipients into the formulations to improve the efficacy of the dosage form. One of the most promising of such excipients, L-leucine, is expected to be approved for inhalation soon and has been studied exhaustively. However, during stability, small fibers protruding from the particles of leucine-containing powders have occasionally been observed. To clarify the origin of these fibers and assess their potential influence on the performance of the powders, three different classes of spray-dried leucine-containing formulation systems were studied over an 8-month accelerated stability program. These systems consisted of a large molecule biologic (bevacizumab) in conjunction with a glass former (trehalose), an amorphous small-molecular mass active (moxidectin), and a crystallizing active (mannitol). It was determined that the appearance of the fibers was due to the presence of small quantities of leucine in higher energy states, either because these were amorphous or present as a less stable crystalline polymorph. It was further shown that the growth of these leucine fibers caused no significant physicochemical instability in the powders. Nor, more importantly, did it decrease their aerosol performance in a dry powder inhaler or reduce the concentration of their active pharmaceutical ingredients.

Lipidoid nanoparticles increase ATP uptake into hypoxic brain endothelial cells

Lipidoid nanoparticles (LNPs) are clinically successful carriers for nucleic acid delivery to liver and muscle targets. The ability of LNPs to load and deliver small molecule drugs has not been reported yet. We propose that the delivery of adenosine triphosphate (ATP) to brain endothelial cells (BECs) lining the blood–brain barrier may increase cellular energetics of the injured BECs. We formulated and studied the physicochemical characteristics of ATP-loaded LNPs using the C12-200 ionizable cationic lipid and other helper lipids. Polyethylene glycol-dimyristoyl glycerol (PEG-DMG), one of the helper lipids, played a crucial role in maintaining colloidal stability of LNPs over time whereas the inclusion of both ATP and PEG-DMG maintained the colloidal stability of LNPs in the presence of serum proteins. ATP-LNPs formulated with PEG-DMG resulted in a 7.7- and 6.6- fold increased uptake of ATP into normoxic and hypoxic BECs, respectively. Altogether, our results demonstrate the potential of LNPs as a novel carrier for the delivery of small molecular mass actives to BECs—a CNS target.

Enhanced three-dimensional electrochemical process using magnetic recoverable of Fe3O4@GAC towards furfural degradation and mineralization

Furfural is as one of the major environmental pollutants in different industrial effluents such as refinery and petrochemical, paper, cardboard and oil refining. This toxic chemical is irritant and causes allergy for skin, eyes and mucous membranes. This study was developed to investigate the efficiency of a three-dimensional electrochemical process in the presence of granular activated carbon magnetized with Fe 3 O 4 (Fe 3 O 4 @GAC) particle electrodes for removal of furfural from aqueous solution. The particle electrodes structural and morphological featured were determined via BET, VSM, XRD, FE-SEM and FTIR techniques. The experiments were performed based on central composite design (CCD) and the role of influencing factors including reaction time, pH, voltage and initial furfural concentrations at five levels were evaluated. The Quadratic model with high correlation coefficient = 0.9872 (R 2 and (R 2 Adj = 0.9724)) was suggested for experimental data analysis. The performance of electrochemical oxidation towards furfural degradation was enhanced substantially after adding Fe 3 O 4 @GAC. The highest furfural removal efficiency (98.2%) was achieved under optimal conditions (furfural: 201 mg/L, electrolysis time: 69 min, voltage: 19 V, and pH: 5.0). Besides, over 78 and 74 % of COD and TOC were removed by Fe 3 O 4 @GAC-based three-dimensional process, respectively. Based on the COD/TOC ratio and average oxidation state (AOS) index, a significant increase was observed in the biodegradability of intermediates of furfural after treatment. Results showed that three-dimensional electrochemical process with particle electrodes is a promising technology for efficient removal of furfural, even at high concentrations. Results of Liquid chromatography–mass spectrometry (LC-MS) analysis and degradation pathway showed that furfural could be oxidized to compounds with smaller molecular masses, which eventually converted to carbon dioxide and water.

Melianone inhibits Secreted Aspartic Proteases (SAP), a Virulence Factor During Hyphal Formation in Candida albicans.

Candida albicans (C.-P. Robin) Berkhout, the pathogenic yeasts' ability to transform from yeast to hyphal forms in the bloodstream is essential during systemic infections. Among the several virulence factors studied, secreted aspartic proteinases (SAPs) involved in hyphal penetration are targets of putative hyphal inhibitors. Upregulation of SAP6 gene, (two-to 31- fold high) during budded to hyphal transition and lack of studies on its inhibition, prompted us to investigate this particular protein using in silico tools. Hyphal inhibition of germinating yeast cells by melianone, a triterpenoid, from Swietenia mahagoni (L.) Jacq. (Meliaceae) was observed at 0.1 μM (IC50). One of the targets of putative hyphal inhibitors, SAP, was assayed and for the first time, 50 % of the biological SAP activity was found to be inhibited by melianone at 0.125 μM. This data on SAP inhibition led us to analyse the 3-dimensional structure for SAP6 protein that was constructed through a combination of homology modelling and ab-initio method (Phyre2) and validated before performing Induced Fit Docking (IFD). Melianone formed H-bond and hydrophobic interactions with the crucial residues (ASP108, TYR160, ALA161, ASP162, ASP294, THR297, ASP379) in the catalytic site of SAP6 with a glide energy (-)54.9327 kcal/mol upon Induced Fit Docking (IFD). We report here for the first time on the SAP inhibitory ability of melianone at 0.125 uM. Being a small molecular mass inhibitor, binding with high affinity to the S3 pocket sites of SAP proteins provides evidence for pre-clinical testing of such compounds against fungal pathogens. The study is a valuable insight for further research on novel and effective inhibitors targeting SAP.

Bacteria binding and antibacterial activities of three small heat shock protein from the swimming crab, Portunus trituberculatus

Small heat shock proteins (sHSPs), a group of molecular chaperones with small molecular mass, are involved in immune responses in many organisms. However, the actual roles of sHSPs in immune defence against bacteria remain largely unclear. In this study, three sHSPs, named PtHSP10, PtHSP20 and PtHSP40 were identified from Portunus trituberculatus, and their immune function was investigated. Sequence analysis showed that the deduced amino acid sequences of PtHSP10, PtHSP20 and PtHSP40 exhibited conserved structures and the functional features. The tissue distributions and transcriptional profiles of these sHSPs in response to Vibrio alginolyticus challenge were analysed by quantitative real-time PCR (qRT-PCR). The results showed that the genes of the three sHSPs were ubiquitously expressed in various tissues, and mainly expressed in hepatopancreas or muscle. The transcriptions of these sHSPs in hepatopancreas of P. trituberculatus injected with V. alginolyticus were upregulated rapidly at 3 h post injection. Moreover, bacterial binding assay showed that the three sHSPs displayed distinct binding activities towards Gram-negative bacteria (V. alginolyticus and V. parahaemolyticus) and Gram-positive bacteria (Staphylococcus aureus). PtHSP10 could bind to V. parahaemolyticus and S. aureus, while PtHSP40 exhibited binding activity towards V. alginolyticus and V. parahaemolyticus. No binding was observed between PtHSP20 and the three tested bacteria. Moreover, PtHSP40 showed antibacterial activity against V. alginolyticus. Our findings indicated that these sHSPs displayed different immune function in immune defence.

Hydrogen, a Novel Therapeutic Molecule, Regulates Oxidative Stress, Inflammation, and Apoptosis.

Molecular hydrogen (H2) is a colorless and odorless gas. Studies have shown that H2 inhalation has the therapeutic effects in many animal studies and clinical trials, and its application is recommended in the novel coronavirus pneumonia treatment guidelines in China recently. H2 has a relatively small molecular mass, which helps it quickly spread and penetrate cell membranes to exert a wide range of biological effects. It may play a role in the treatment and prevention of a variety of acute and chronic inflammatory diseases, such as acute pancreatitis, sepsis, respiratory disease, ischemia reperfusion injury diseases, autoimmunity diseases, etc.. H2 is primarily administered via inhalation, drinking H2-rich water, or injection of H2 saline. It may participate in the anti-inflammatory and antioxidant activity (mitochondrial energy metabolism), immune system regulation, and cell death (apoptosis, autophagy, and pyroptosis) through annihilating excess reactive oxygen species production and modulating nuclear transcription factor. However, the underlying mechanism of H2 has not yet been fully revealed. Owing to its safety and potential efficacy, H2 has a promising potential for clinical use against many diseases. This review will demonstrate the role of H2 in antioxidative, anti-inflammatory, and antiapoptotic effects and its underlying mechanism, particularly in coronavirus disease-2019 (COVID-19), providing strategies for the medical application of H2 for various diseases.

Solvent-free synthesis of nitrone-containing template as a chemosensor for selective detection of Cu(II) in water

A state-of-the-art method was developed for repurposing nitrone-containing compounds in the chemosensory field, the ability of the designed molecules to chelate metal cations was evaluated, and their unprecedented solubility in water was confirmed. A facile, rapid, and solvent-free method of synthesizing small molecular mass chemosensors was developed by using a modulative α-aryl-N-aryl nitrone template. α-(Z)-Imidazol-4-ylmethylen-N-phenyl nitrone (Nit1) and α-(Z)-2-pyridyl-N-phenyl nitrone (Nit2) were prepared in 15 min, isolated in less than 60 min with ca. 90% yield, and screened against nine metal cations. Nit1 is a small-molecular-mass compound (188 g mol−1) that is water-soluble and has specificity for sensing Cu2+ with an association constant of K = 1.53 × 1010 and a limit of detection (LOD) of 0.06 ppm. These properties make Nit1 a competitive chemosensor for the detection of Cu2+ in aqueous solution. The nitrone-containing template used in this study is a step forward for new and small chemosensory entities.

Sub-Micromolar Inhibition of SARS-CoV-2 3CLpro by Natural Compounds

Inhibiting the main protease 3CLpro is the most common strategy in the search for antiviral drugs to fight the infection from SARS-CoV-2. We report that the natural compound eugenol is able to hamper in vitro the enzymatic activity of 3CLpro, the SARS-CoV-2 main protease, with an inhibition constant in the sub-micromolar range (Ki = 0.81 μM). Two phenylpropene analogs were also tested: the same effect was observed for estragole with a lower potency (Ki = 4.1 μM), whereas anethole was less active. The binding efficiency index of these compounds is remarkably favorable due also to their small molecular mass (MW < 165 Da). We envision that nanomolar inhibition of 3CLpro is widely accessible within the chemical space of simple natural compounds.

Research Status of Antigen Synthesis Methods

Most haptens are small molecules compounds. Because of its small molecular mass itself alone can not induce an immune response, it is necessary to form combined with the carrier hapten-carrier protein conjugate can stimulate the body's immune response. In recent years, in-depth research has been conducted at home and abroad on the synthesis of artificial antigens and the analysis of the results. Artificial antigen synthesis technology has been applied to many fields such as food and environment, which have more or less impact on our lives. This article reviews how to select carrier protein and the coupling method of hapten and carrier in the process of artificial antigen synthesis.