New human single chain anti-idiotypic antibody against benzo[a]pyrene
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 specif...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Termedia Publishing House
2017-08-01
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| Series: | Central European Journal of Immunology |
| Subjects: | |
| Online Access: | https://www.termedia.pl/New-human-single-chain-anti-idiotypic-antibody-against-benzo-a-pyrene,10,30439,1,1.html |
| Summary: | 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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| ISSN: | 1426-3912 1644-4124 |