New human single chain anti-idiotypic antibody against benzo[a]pyrene
| Autor: | Artem E. Studennikov, Valentin A. Ustinov, Vera V. Morozova, Nina V. Tikunova, Andrey N. Glushkov |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: | |
| Zdroj: | Central European Journal of Immunology, Vol 42, Iss 2, Pp 123-130 (2017) |
| Druh dokumentu: | article |
| ISSN: | 1426-3912 1644-4124 |
| DOI: | 10.5114/ceji.2017.69353 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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