EXPERIMENTAL IMMUNOLOGY
New human single chain anti-idiotypic antibody against benzo[a]pyrene
Submission date: 2016-06-06
Final revision date: 2016-10-01
Acceptance date: 2016-10-01
Publication date: 2017-08-08
Cent Eur J Immunol 2017;42(2):123-130
KEYWORDS
anti-idiotypic antibodybenzo[a]pyreneimmunologyphage displaypolycyclic aromatic hydrocarbonssingle-chain antibody
ABSTRACT
The naïve library from the lymphocytes of healthy humans was screened by murine single-stranded idiotypic antibodies against benzo[a]pyrene (pSh). The phage clone which contained of anti-idiotypic antibody against benzo[a]pyrene, designated as A4, was chosen for further work because of highly specific to pSh. The available protein databases were searched. The A4 amino acid sequence was unique and 76% identical to a sequence in antibody against interferon g. The A4 protein was expressed in bacteria and purified by two different methods: His-tagged A4 and CBD-fusion A4. Both the A4 bound to pSh and also to the human single chain idiotypic antibody against the benzo[a]pyrene (T72) by ELISA. The Kd values of A4 for pSh and T72 were very close: 4.44 × 10-7 M and 5.71 × 10-7M, respectively. A4 was a competitor with benzo[a]pyrene for binding sites of both idiotypic pSh and T72 in competitive ELISA. Thus, A4 was a high affinity anti-idiotypic against benzo[a]pyrene which recognised pSh and T72 active sites.
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| eISSN: | 1644-4124 |
| ISSN: | 1426-3912 |
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