Synthesis Of Artificial Antigen Research Articles

Artificial antibody-antigen-directed immobilization of α-amylase to hydrolyze starch for cascade reduction of 2-nitro-4-methylphenol to 2-amino-4-methylphenol

Nitrophenol is a hazardous substance that poses a threat to the environment and human health, and its treatment has attracted widespread attention. The purpose of this study is to establish an environmentally friendly α-amylase system for the hydrolysis of starch to reduce nitrophenol to aminophenol through cascade reactions. The α-amylase system was obtained through artificial antibody-antigen-directed immobilization, including the synthesis of artificial antibodies, synthesis of artificial antigens, and affinity assembly. In this process, catechol and protocatechuic aldehyde were used to prepare artificial antibodies and artificial antigens respectively through polymerization and Schiff base reactions. Then, artificial antibodies captured the catechol in the artificial antigen structure to form immobilized α-amylases. Compared with free α-amylase, the immobilized α-amylase showed a good reusability and excellent regenerative ability. Subsequently, the immobilized α-amylase were used in the reaction of catalyzing starch hydrolysis to synthesize 2-amino-4-methylphenol, and the yield of 2-amino-4-methylphenol was 58.88 ± 0.19 %. After 5 consecutive catalytic reactions, a yield of 47.61 ± 1.27 % can still be achieved.

Biomimetic immobilization of α-glucosidase inspired by antibody-antigen specific recognition for catalytic preparation of 4-methylumbelliferone

Immobilization technology plays an important role in enhancing enzyme stability and environmental adaptability. Despite its rapid development, this technology still encounters many challenges such as enzyme leakage, difficulties in large-scale implementation, and limited reusability. Drawing inspiration from natural paired molecules, this study aimed to establish a method for immobilized α-glucosidase using artificial antibody-antigen interaction. The proposed method consists of three main parts: synthesis of artificial antibodies, synthesis of artificial antigens, and assembly of the artificial antibody-antigen complex. The critical step in this method involves selecting a pair of structurally similar compounds: catechol as a template for preparing artificial antibodies and protocatechualdehyde for modifying the enzyme to create the artificial antigens. By utilizing the same functional groups in these compounds, specific recognition of the antigen by the artificial antibody can be achieved, thereby immobilizing the enzymes. The results demonstrated that the immobilization amount, specific activity, and enzyme activity of the immobilized α-glucosidase were 25.09 ± 0.10 mg/g, 5.71 ± 0.17 U/mgprotein and 143.25 ± 1.71 U/gcarrier, respectively. The immobilized α-glucosidase not only exhibited excellent reusability but also demonstrated remarkable performance in catalyzing the hydrolysis of 4-methylumbelliferyl-α-D-glucopyranoside.

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.

Research Progress in Antigen Synthesis

In recent years, immunoassay using small molecular compound as hapten has been applied in many fields, such as food and drug, environmental protection, etc., which has achieved ideal detection results. Only when the small molecule compound can only be coupled with the carrier to form an artificial antigen, can it indirectly induce B cells to proliferate and differentiate by T cell epitope, and then specific antibodies can be produced. The synthesis of efficient artificial antigen is the premise and key to immunoassay. This essay is a literature review at home and abroad about the design and synthesis methods of small molecular haptens, the selection of carriers, the coupling methods of haptens and carriers, the purification and identification methods, which are involved in the synthesis of artificial antigens.

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.

Progress in the Synthesis of Artificial Antigen

Progress in the Synthesis of Artificial Antigen

Development of two enzyme-linked immunosorbent assay formats for thifluzamide residues’ analysis based on distinct polyclonal antibodies

ABSTRACT Haptens 2-Methyl-4-(trifluoromethyl)thiazole-5-carboxylic acid and 2,6-Dibromo-4-(trifluoromethoxy)aniline, the two moieties of thifluzamide, were conjugated with carrier proteins for the synthesis of artificial antigens. Two distinct anti-thifluzamide polyclonal antibodies (PAb-1 and PAb-2) were produced from the immunized female Balb/c mice. The indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) in two formats based on the PAbs was developed for thifluzamide analysis. The concentration of 50% inhibition (IC50) of ELISA-1 was 1.39 mg L−1 and its limit of detection (LOD) was 0.082 mg L−1. Meanwhile, ELISA-2 had a similar IC50 of 1.96 mg L−1 and a LOD of 0.074 mg L−1 as ELISA-1. Both the raised PAbs exhibited high specificity to thifluzamide. The recoveries for spiked samples including water and wheat ranged from 72.0% to 128.4%, and the accuracy of ELISA was confirmed by high-performance liquid chromatography. In summary, the ic-ELISA might be a promising tool for simple, sensitive and rapid detection of thifluzamide residues in real samples.

Synthesis of artificial antigen and preparation of polyclonal antibody against okadaic acid

Okadaic acid(OA)is one kind of lipophilic marine biotoxins,and main pathogenic factor of diarrheic shellfish poisoning(DSP).In this paper,the artificial antigen was made by coupling hapten of OA with keyhole limpet hemocyanin(KLH)using active ester method,and the artificial antigen was tested by SDS-polyacrylamide gel electrophoresis and IR spectrum.After being obtained from rabbits,the antiserum was purified by Chromatography on Protein A Sepharose CL-4B gel column,and the purification efficiency was verified by ultraviolet scanning and ciELISA.The results showed the artificial antigen was prepared successfully and lots of interfering substances were removed.After being purified,the concentration of antiserum was 2.16 mg/mL,it had high titer(12 800)and sensitivity(the value of IC15 was 3.41 ng/mL),which was tested by ciELISA.The success of obtaining polyclonal antibody against OA with high titer and affinity provides the solid foundation which is practical and theoretical for developing rapid and economical detections.

Recent Progress in Synthetic Methods of Small Molecule Antigen

With increasing applications of immune assay,the synthesis of artificial antigen with good stability and immunogenic is the key and premise to prepare the antibody and to establish the method for small molecule immune assay.The progress of synthesis and design method of hapten,carrier selection and the couple method of hapten with carrier are reviewed in this paper.In addition,some questions of the synthesis of small molecule artificial antigen were also discussed.

Recent Advances in the Synthesis of Artificial Antigen and Its Application in the Detection of Pesticide Residue

Recent advances in the research of artificial antigen have shown that artificial antigens can be valuable approach for the treatment of some diseases as well as the detection of pesticide residues. By directly/indirectly coupling hapten to an appropriated carrier (macromolecule), artificial antigen can induce animals to produce hapten-specific antibody. Based on this principle, various vaccines have been developed. More impotently, new analytical method, immunological analysis has also been established. Comparing the conventional technologies, such as chromatographic methods, this promising method offers an alternative with high specificity, sensitivity, simplicity and suitability for the analysis of a large number of samples in a short period of time. In this review, we describe the recent advances in the synthesis of artificial antigen and its application in the detection of pesticide residues.

Synthesis of fragments of bacterial polysaccharides and their application for the preparation of synthetic antigens

Abstract