POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5
The aim of the research was to obtain the polyclonal antibodies against a fragment of human plasminogen kringle 5 and to study their immunochemical properties. The following approaches were used: immunization of rabbits with plasminogen kringle 5, receiving of high-titer immune serum, synthesis of k...
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| Format: | Article |
| Language: | English |
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2017-06-01
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| Series: | Biotechnologia Acta |
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| Online Access: | http://biotechnology.kiev.ua/images/storage/3_2017/kapustianenko_3_2017.pdf |
| _version_ | 1797728075975229440 |
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| author | Kapustianenko L. G. |
| author_facet | Kapustianenko L. G. |
| author_sort | Kapustianenko L. G. |
| collection | DOAJ |
| description | The aim of the research was to obtain the polyclonal antibodies against a fragment of human plasminogen kringle 5 and to study their immunochemical properties. The following approaches were used: immunization of rabbits with plasminogen kringle 5, receiving of high-titer immune serum, synthesis of kringle 5-based affinity sorbent for selection of monospecific antibodies, chromatography on synthesized K 5-Sepharose, ELISA immunoblot assay.
The obtained polyclonal antibodies reacted in ELISA with a plasminogen K 5 fragment, and to a much lesser extent with mini-plasminogen, Lys-plasminogen, K 1-3 and K 4 plasminogen fragments, Glu-plasminogen, in descending order. Based on the dissociation constant determination it was found that these antibodies had high affinity to their epitopes within K 5 fragment (3.89・10–10 M), mini-plasminogen (5.46・10–9 М) and Lys-plasminogen (2.54・10–9 М), and insignificant affinity to K1-3, K4 and Glu-plasminogen. These antibodies reacted in immunoblotting assay with an isolated K 5 fragment of human plasminogen, Lys-Pg and mini-Pg, and did not react with K 5 in Glu-Pg and with K 1-3 or K 4 fragments. Thus, the obtained polyclonal antibodies were monospecific and had a high affinity to kringle 5. These antibodies can be used for the development immuno chemical and immunosensory methods for the quantitative determination of angiostatin K 5 in biological materials. |
| first_indexed | 2024-03-12T11:08:38Z |
| format | Article |
| id | doaj.art-7735a0fc43f241378a6c815759ac3d86 |
| institution | Directory Open Access Journal |
| issn | 2410-7751 2410-776X |
| language | English |
| last_indexed | 2024-03-12T11:08:38Z |
| publishDate | 2017-06-01 |
| publisher | National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
| record_format | Article |
| series | Biotechnologia Acta |
| spelling | doaj.art-7735a0fc43f241378a6c815759ac3d862023-09-02T03:23:49ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryBiotechnologia Acta2410-77512410-776X2017-06-01103414910.15407/biotech10.03.041POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5Kapustianenko L. G. 0Palladian Institute of Biochemistry of the National Academy of Sciences of Ukraine, KyivThe aim of the research was to obtain the polyclonal antibodies against a fragment of human plasminogen kringle 5 and to study their immunochemical properties. The following approaches were used: immunization of rabbits with plasminogen kringle 5, receiving of high-titer immune serum, synthesis of kringle 5-based affinity sorbent for selection of monospecific antibodies, chromatography on synthesized K 5-Sepharose, ELISA immunoblot assay. The obtained polyclonal antibodies reacted in ELISA with a plasminogen K 5 fragment, and to a much lesser extent with mini-plasminogen, Lys-plasminogen, K 1-3 and K 4 plasminogen fragments, Glu-plasminogen, in descending order. Based on the dissociation constant determination it was found that these antibodies had high affinity to their epitopes within K 5 fragment (3.89・10–10 M), mini-plasminogen (5.46・10–9 М) and Lys-plasminogen (2.54・10–9 М), and insignificant affinity to K1-3, K4 and Glu-plasminogen. These antibodies reacted in immunoblotting assay with an isolated K 5 fragment of human plasminogen, Lys-Pg and mini-Pg, and did not react with K 5 in Glu-Pg and with K 1-3 or K 4 fragments. Thus, the obtained polyclonal antibodies were monospecific and had a high affinity to kringle 5. These antibodies can be used for the development immuno chemical and immunosensory methods for the quantitative determination of angiostatin K 5 in biological materials.http://biotechnology.kiev.ua/images/storage/3_2017/kapustianenko_3_2017.pdfhuman plasminogen fragmentskringle 5polyclonal antibodiesangiostatines |
| spellingShingle | Kapustianenko L. G. POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 Biotechnologia Acta human plasminogen fragments kringle 5 polyclonal antibodies angiostatines |
| title | POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 |
| title_full | POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 |
| title_fullStr | POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 |
| title_full_unstemmed | POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 |
| title_short | POLYCLONAL ANTIBODIES AGAINST HUMAN PLASMINOGEN KRINGLE 5 |
| title_sort | polyclonal antibodies against human plasminogen kringle 5 |
| topic | human plasminogen fragments kringle 5 polyclonal antibodies angiostatines |
| url | http://biotechnology.kiev.ua/images/storage/3_2017/kapustianenko_3_2017.pdf |
| work_keys_str_mv | AT kapustianenkolg polyclonalantibodiesagainsthumanplasminogenkringle5 |