Peculiarities of interaction of monoclonal antibody B2 with polycyclic aromatic hydrocarbons and peptide-mimotope of benzo[a]pyrene

Глушков, А. Н.; Apalko, Svetlana V.; Bakulina, Anastasia; Матвеева, В. А.; Khrapov, E. A.; Kostyanko, M. V.; Silnikov, Vladimir N.; Филипенко, М. Л.

doi:10.1134/s0026893310040175

Cited by 4 publications

(12 citation statements)

References 16 publications

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“…We have previously shown mAb B2 high affinity interaction with a group of PAHs: BP, Ac, Ba, Cr, and Py. [20] For pSh-CBD the highest binding was identified to Ac ( Figure 4). For other tested PAHs interaction was lower and almost equal.…”

Section: Binding Psh-cbd With Pahsmentioning

confidence: 99%

“…[20] According to this model pSh-CBD there is a pocket for binding of BP. To confirm the hypothetical aa involved in the coordination of BP, we mutated corresponding aa to Ala, in groups and individually.…”

Section: Determination Of Aa Psh-cbd In Bp-linkingmentioning

confidence: 99%

“…[20] Figure 1 shows the aa sequence analysis of pSh and comparison with published heavy and light chains of Ab against BP. [19] Comparison pSh aa sequences with published mouse antibody showed 38%, 39%, and 55% identity to the heavy chains (KpH1, KpH2, and KpH3, respectively) and 60%, 58%, and 84% for light chains (KpL1, KpL2, and KpL3, respectively).…”

Section: Amino Acid Sequence Analysis Of Protein Pshmentioning

confidence: 99%

“…[19] We cloned heavy and light chains of mAb named B2 from hybridoma cells. [20] The individual variable domains of the Ab were assembled into a single structure, as has been done in our previous article. [20] The present article was aimed at scFv purifying, characterization, and comparison with mAb B2, effectively linking PAHs.…”

mentioning

confidence: 99%

“…[20] The individual variable domains of the Ab were assembled into a single structure, as has been done in our previous article. [20] The present article was aimed at scFv purifying, characterization, and comparison with mAb B2, effectively linking PAHs. This Ab can subsequently serve as binding subunit in bioengineering, and to produce test systems to determine the BP by Ab.…”

mentioning

confidence: 99%

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Purification and Characterization of Mouse Single-Chain Antibody against Polycyclic Aromatic Hydrocarbons

Ustinov¹,

Averjanov²,

Глушков³

2014

Journal of Immunoassay and Immunochemistry

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Polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyren mainly induce lung cancer in humans. We characterized the mouse single chain antibody against benzo[a]pyren (pSh). pSh was expressed and purified as cellulose binding domain fusion (pSh-CBD). The pSh-CBD bound five different PAH with high affinity. The 18 amino acid linker connected pSh-CBD heavy and light chains provided correct protein folding. The KDs for pSh-CBD and polycyclic aromatic hydrocarbons were similar to KDs for monoclonal antibody, approximately 10(-8). Separately heavy and light chains of pSh-CBD did not interact with benzo[a]pyren. Previously defined eleven pSh-CBD aa involved to benzo[a]pyren binding were confirmed by mutagenesis.

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Section: Binding Psh-cbd With Pahsmentioning

confidence: 99%

Section: Determination Of Aa Psh-cbd In Bp-linkingmentioning

confidence: 99%

Section: Amino Acid Sequence Analysis Of Protein Pshmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Purification and Characterization of Mouse Single-Chain Antibody against Polycyclic Aromatic Hydrocarbons

Ustinov¹,

Averjanov²,

Глушков³

2014

Journal of Immunoassay and Immunochemistry

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Idiotypic and anti-idiotypic antibodies against polycyclic aromatic hydrocarbon in human blood serum are new biomarkers of lung cancer

et al. 2019

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Evaluation of epidemiologic risk factor in relation to lung cancer invoked by polycyclic aromatic hydrocarbons has been inconsistent. To address this issue, we conducted a prospective evaluation of new biomarkers for lung cancer classified according levels of idiotypic and anti-idiotypic antibodies against polycyclic aromatic hydrocarbons in human blood serum. The blood serums of 557 lung cancer patients and 227 healthy donors were analysis of these antibodies by ELISA. Collected data were regrouped and analyzed by gender, smoking, and age as predictors of risk lung cancer factors. Also, the data of lung cancer patients were additionally analyzed by stages and types of lung cancer, surgery, and chemotherapy. It was suggested to use ratio of idiotypic and anti-idiotypic antibodies rather than distinguish level each of them separately. The ratio of levels in healthy people was 3.32 times higher than in lung cancer patients. This approach gave more precisely results and great prognostic value. The logistic regression model (AUC = 0.9) and neural networks (AUC = 0.95) were built to compare lung cancer patients and healthy donors by predictors. The ELISA data of 49 people random sampled from the originally ELISA data and ELISA data of 52 coal miners as a group of lung cancer risk were confirmed logistic regression model. So, suggested idiotypic and anti-idiotypic antibodies against polycyclic aromatic hydrocarbons were not only shown difference between healthy donors and lung cancer patients also elicited group of lung cancer risk among healthy people.

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Generation and Characterization of Human Single-Chain Antibodies Against Polycyclic Aromatic Hydrocarbons

Ustinov¹,

Studennikov²,

Vavilov³

et al. 2015

Immunological Investigations

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Polycyclic aromatic hydrocarbons (PAHs) are widely distributed and relocated in the environment as a result of the incomplete combustion of organic matter. Many PAHs and their epoxides are highly toxic, mutagenic and/or carcinogenic to microorganisms as well as to higher systems including humans. BP is one of the most toxicologically active PAHs and is often used as a prototype for this entire class of contaminants. In order to select anti-BP antibodies, the conjugate of BP with BSA (BP-BSA) was used to screen naïve combinatorial phage library of human scFvs. Seven unique scFvs against BP-BSA were selected after three rounds of selection. Analysis of the genes encoding the scFvs subdivided them to gene families and subfamilies. Homology with the closest germline ranged from 80.21% to 97.57% for heavy chains and 88.89% to 98.57% for the light chains. Four of the seven scFv amino acid residues sequences without stop codons in frame were selected for proteomic analysis with each other. Four scFvs encoded unique non-related proteins with low-sequence identity among them. All CDRs and the boundaries in the CDR3 formation were carried out. Two of the scFvs (T68 and T72) with the highest binding capabilities to PAHs were expressed in E. coli and purified using a nickel resin. The KDs of T68 to BP-BSA, chrysene, pyrene, and benzo[a]anthracene were almost similar, approximately 10(-7 )M. The KDs of T72 to benzo[a]anthracene and chrysene were 9.42 × 10(-8 )M and 2.63 × 10(-7 )M, respectively. The computational models of T68 and T72 active centers were different.

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Peculiarities of interaction of monoclonal antibody B2 with polycyclic aromatic hydrocarbons and peptide-mimotope of benzo[a]pyrene

Cited by 4 publications

References 16 publications

Purification and Characterization of Mouse Single-Chain Antibody against Polycyclic Aromatic Hydrocarbons

Purification and Characterization of Mouse Single-Chain Antibody against Polycyclic Aromatic Hydrocarbons

Idiotypic and anti-idiotypic antibodies against polycyclic aromatic hydrocarbon in human blood serum are new biomarkers of lung cancer

Generation and Characterization of Human Single-Chain Antibodies Against Polycyclic Aromatic Hydrocarbons

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