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

Won Ki Min; Dae Hyuk Kweon; Kyungmoon Park; Yong Cheol Park; Jin Ho Seo

doi:10.1016/j.foodchem.2010.11.088

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

Won Ki Min, Dae Hyuk Kweon, Kyungmoon Park, Yong Cheol Park, Jin Ho Seo

Department of Integrative Biotechnology

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	1316-1323
Number of pages	8
Journal	Food Chemistry
Volume	126
Issue number	3
DOIs	https://doi.org/10.1016/j.foodchem.2010.11.088
State	Published - 1 Jun 2011

Keywords

Aflatoxin B
Escherichia coli
Hybridoma cell
Monoclonal antibody
Single-chain variable fragment

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

abstract = "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 (KD), which was about 17 times higher than the parental anti-AFB1 mAb of 6.95 × 10-9 M.",

keywords = "Aflatoxin B, Escherichia coli, Hybridoma cell, Monoclonal antibody, Single-chain variable fragment",

author = "Min, \{Won Ki\} and Kweon, \{Dae Hyuk\} and Kyungmoon Park and Park, \{Yong Cheol\} and Seo, \{Jin Ho\}",

year = "2011",

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doi = "10.1016/j.foodchem.2010.11.088",

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T1 - Characterisation of monoclonal antibody against aflatoxin B1 produced in hybridoma 2C12 and its single-chain variable fragment expressed in recombinant Escherichia coli

AU - Min, Won Ki

AU - Kweon, Dae Hyuk

AU - Park, Kyungmoon

AU - Park, Yong Cheol

AU - Seo, Jin Ho

PY - 2011/6/1

Y1 - 2011/6/1

N2 - 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 (KD), which was about 17 times higher than the parental anti-AFB1 mAb of 6.95 × 10-9 M.

AB - 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 (KD), which was about 17 times higher than the parental anti-AFB1 mAb of 6.95 × 10-9 M.

KW - Aflatoxin B

KW - Escherichia coli

KW - Hybridoma cell

KW - Monoclonal antibody

KW - Single-chain variable fragment

UR - https://www.scopus.com/pages/publications/78751572849

U2 - 10.1016/j.foodchem.2010.11.088

DO - 10.1016/j.foodchem.2010.11.088

M3 - Article

AN - SCOPUS:78751572849

SN - 0308-8146

VL - 126

SP - 1316

EP - 1323

JO - Food Chemistry

JF - Food Chemistry

IS - 3

ER -

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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Keywords

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