Characterisation of monoclonal antibody against aflatoxin B₁ produced in hybridoma 2C12 and its single-chain variable fragment expressed in recombinant Escherichia coli

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• 作者：Won-Ki Min^a ; Dae-Hyuk Kweon^b ; Kyungmoon Park^c ; Yong-Cheol Park^d ; ^{ycpark@kookmin.ac.kr} ; Jin-Ho Seo^a ; ^{jhseo94@snu.ac.kr}
• 关键词：Aflatoxin B₁ ; Monoclonal antibody ; Single-chain variable fragment ; Hybridoma cell ; Escherichia coli
• 刊名：Food Chemistry
• 出版年：2011
• 出版时间：1 June 2011
• 年：2011
• 卷：126
• 期：3
• 页码：1316-1323
• 全文大小：624 K

文摘

An anti-aflatoxin B₁ monoclonal antibody (anti-AFB₁ mAb) from the hybridoma 2C12 was established and its inhibition concentration fifty (IC₅₀) for AFB₁ and relative cross-reactivities (CRs) to other mycotoxins were estimated to be 8 ng/mL and less than 4 % compared with AFB₁ by a competitive direct enzyme-linked immunosorbent assay. For production of anti-AFB₁ single-chain variable fragment (anti-AFB₁ scFv) in recombinant Escherichia coli, its scFv-coding genes were cloned from the hybridoma 2C12. The anti-AFB₁ scFv formed inclusion bodies in the cytoplasm of E. coli required in vitro refolding process and hence recovered to retain binding activity successfully. Surface plasmon resonance analysis resulted that anti-AFB₁ scFv possessed 1.16 × 10⁻⁷ M of equilibrium dissociation constant (K_D), which was about 17 times higher than the parental anti-AFB₁ mAb of 6.95 × 10⁻⁹ M.