CN-116333106-B - Aflatoxin B1Antibodies and uses

Abstract

The invention provides an aflatoxin B1 antibody and application thereof, and in particular relates to an aflatoxin B1 antibody only comprising a heavy chain variable region, nucleotide for encoding the antibody, and an expression vector carrying the aflatoxin B 1 antibody or a nucleotide sequence for encoding the same. The invention screens and prepares the antibody aiming at aflatoxin B 1 from phage nanometer antibody synthesis library and is applied to detection or identification of aflatoxin B 1 . The affinity constant of the antibody is 4.19E-09, and the antibody can effectively detect aflatoxin B 1 and identify other common mycotoxins.

Inventors

• XU QINGQIANG
• MO FENGFENG
• XIAO KAI
• Mao Guanchao
• SUN MINGXUE
• PEI ZHIPENG
• MENG WENQI
• CEN JINFENG
• ZHANG XINKANG

Assignees

• 中国人民解放军海军军医大学

Dates

Publication Date

20260512

Application Date

20220713

Claims (6)

1. 1. The aflatoxin B 1 antibody is characterized in that the antibody is a monoclonal antibody, and the monoclonal antibody contains and only contains a heavy chain variable region, and the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 1.
2. 2. A polynucleotide encoding an aflatoxin B 1 antibody as claimed in claim 1, wherein the polynucleotide sequence is shown in SEQ ID No. 2.
3. 3. An expression vector carrying the polynucleotide sequence encoding the aflatoxin B 1 antibody of claim 2.
4. 4. Use of the aflatoxin B 1 antibody of claim 1 in the preparation of an aflatoxin B 1 detection or identification reagent or kit.
5. 5. Use according to claim 4, characterized in that: Wherein the detection or identification reagent comprises a biomarker or chemically-labeled aflatoxin B 1 antibody; The kit comprises a labeled compound obtained by biomarker or chemical labeling of an aflatoxin B 1 antibody.
6. 6. An aflatoxin B 1 detection or identification kit, comprising a blood sample processing reagent and a labeled complex obtained by biomarker or chemical labeling of the aflatoxin B 1 antibody according to claim 1.

Description

Aflatoxin B 1 antibody and application thereof Technical Field The invention relates to the technical field of biological medicines, relates to an aflatoxin B 1 antibody, and in particular relates to an aflatoxin B 1 specific monoclonal antibody and application thereof. Background Biotoxins are metabolites produced in nature by various organisms such as animals, plants, microorganisms, and the like, and are of a wide variety, mainly proteins, polypeptides, or small molecule compounds. The most common biotoxin is marine paralytic shellfish poisoning and various mycotoxins are small molecular compounds (molecular weight is less than 1 kDa), and the biotoxin is severe in toxicity and wide in source, so that great economic loss is brought to industries such as planting, livestock raising, aquatic products and the like, and harm is also brought to human health and safety. Therefore, the biological toxin detection and treatment research has important significance. Aflatoxin B 1（aflatoxin B1,AFB1) is a secondary metabolite produced by virulent strains such as aspergillus flavus (Aspergillus flavus) and aspergillus parasiticus (Aparasiticus), and is widely used in mildewed grains and products thereof. AFB 1 has stable physicochemical properties, strong toxicity, carcinogenicity, teratogenicity and mutagenicity, and is the most toxic mycotoxin and extremely outstanding harm to human health. AFB 1 has hepatophilia in vivo, and is classified as a class I carcinogen by the world health organization as two most important risk factors for hepatitis B virus infection and liver cancer. AFB 1 is very widely contaminated, and despite the wide variety of chemical mildewcides, the mildewcide effect is not very desirable and is potentially toxic to many humans and animals. At present, no specific therapeutic drug is available for AFB 1 poisoning, and liver protection, toxin expelling treatment and symptomatic treatment can only be performed. Therefore, it is important to prevent AFB 1 poisoning by detecting AFB 1. In recent years, phage antibody display technology has been developed rapidly, and the biotoxin antibody yield has been greatly improved. The antibody libraries established so far comprise three classes, natural antibody libraries, immune antibody libraries and synthetic or semisynthetic antibody libraries. The immune antibody library has strong specificity, but still needs animal immune process, and the immune antibody library and the antibody in the natural antibody library still need further humanized transformation to be applied to the treatment of biotoxin poisoning. The synthetic antibody library is artificially synthesized according to the antibody sequence, so that the humanized reconstruction process is avoided, the death failure risk caused by immunization of animals by biotoxin is avoided, the diversity of antibodies in the library is high, and the synthetic antibody library becomes an important means for developing the biotoxin antibody. The aflatoxin B 1 specific recognition antibody has extremely low yield, the obtained antibody has strong cross property, the specific recognition antibody is difficult to obtain in a natural antibody library and an immune antibody library, and a related method for obtaining the specific antibody by artificial synthesis is also lacking. Disclosure of Invention The invention aims to develop a monoclonal antibody for specifically recognizing aflatoxin B 1, screen aflatoxin B 1 by utilizing a phage nanobody total synthesis library, and design experiments to obtain the specific antibody. Firstly designing a phage nanobody artificial synthesis library, secondly obtaining positive monoclonal specifically recognizing aflatoxin B 1 through screening, and finally verifying the affinity and the cross property of the monoclonal antibody. In a first aspect of the present invention, there is provided an aflatoxin B 1 antibody which is a monoclonal antibody, comprising and consisting only of a heavy chain variable region having the amino acid sequence shown below: EVQLVESGGGLVQPGGSLRLSCAASGRIFSSNVMGWFRQAPGKGRELVAAITWRDGSTYYPDSVEGRFTISRDNAKRMVYLQMNSLRAEDTAVYYCAAGRYTWAASAYNYWGQGTQVTVSS（SEQ ID NO.1）. in a second aspect of the present invention, there is provided a nucleotide encoding the above aflatoxin B 1 antibody, for encoding a heavy chain variable region, the nucleotide sequence of which is as follows: GAGGTGCAGCTGGTGGAGAGCGGTGGTGGTCTCGTGCAGCCCGGCGGTAGTCTGCGCCTCAGCTGTGCCGCCAGCGGTCGCATCTTTAGCAGCAATGTGATGGGCTGGTTTCGCCAAGCCCCCGGCAAAGGTCGCGAACTGGTGGCCGCCATTACCTGGCGCGATGGCAGCACCTATTACCCAGATAGCGTGGAAGGCCGCTTCACCATCAGCCGCGATAACGCCAAGCGCATGGTGTATCTGCAGATGAACAGTCTGCGCGCCGAGGACACCGCCGTGTATTATTGTGCCGCCGGCCGCTATACCTGGGCCGCCAGCGCCTACAATTACTGGGGCCAAGGCACCCAAGTGACCGTGAGCAGC（SEQ ID NO.2）. in a third aspect of the present invention, there is provided an expression vector carrying the above aflatoxin B 1 antibody or nucleotide sequence encoding it. In a fourth aspect, the invention provides the use of