Preparation of Differentiated Recombinant Human β2-Microglobulin and Mouse β2-Microglobulin Proteins for its Detection in Class I HLA Chimeric Molecules

Author:

Karkischenko V. N.1,Ezerskiy V. A.2,Koloskova E. M.2,Nesterov M. S.1

Affiliation:

1. Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia

2. All-Russian Research Institute of Physiology, Biochemistry and Animal Nutrition — Branch of the Federal Scientific Centre of Animal Husbandry — All-Russian Institute of Animal Husbandry named after Acad. L.K. Ernst

Abstract

Transgenic humanized animals are increasingly in demand for biomedical research and pharmacological testing. More and more lines of transgenic animals are being created, including those with knockout of their own genes. There is an urgent need for an evidence base for the integration of a transgene, its expression, determination of the knockout state of its own gene at the molecular genetic level, detection of translation of the target protein in different organs and tissues, proof for the absence of protein synthesis (or its non-functionality), the gene of which has been modified. This requires highly specific reagents, proteins and antibodies to them in particular, the vast majority of which are presented by foreign manufacturers. The task was set to identify mouse and human β2-microglobulin in protein fractions of organs and tissues of transgenic and knockout mice of several HLA lines created in recent years at the Scientific Center of Biomedical Technologies, Russia. At the first stage of our research, recombinant E. coli producing strains were obtained.

Publisher

Scientific Center for Biomedical Technologies of the Federal Medical-Biological Agency

Reference15 articles.

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2. Karkischenko N.N., Lazarev V.N., Manuvera V.A., Bobrovsky P.A., Petrova N.V., Koloskova E.M., Glotova E.S. Principy sozdaniya genno-inzhenernoj konstrukcii dlya polucheniya gumanizirovannyh transgennyh myshej, nesushchih gen HLA-C*07:02:01:01, kak proobraz innovacionnyh transgenno-nokautnyh biomodelej [Principles of Creation of a Genetic Engineering Construction for Obtaining Humanized Transgenic Mice with HLA-C*07:02:01:01, as a Promote of Innovative Transgenic and Knockout Biomodels]. Biomedicina [Journal Biomed]. 2024;20(1):8–20. (In Russian). DOI: 10.33647/2074-5982-20-1-8-20.

3. Karkischenko V.N., Petrova N.V., Savchenko E.S., Ogneva N.S., Koloskova E.M., Maksimenko S.V., Manuvera V.A., Bobrovsky P.A., Lazarev V.N. Sozdanie polnih gibritnih DNKkonstrukcij s genom cheloveka HLA-A*02:01:01:01 [Chimeric Construct Engineering with Human Variant HLA-A*02:01:01:01]. Biomedicina [ Journal Biomed]. 2021;17(1):10–23. (In Russian). DOI: 10.33647/2074-5982-17-1-10-23.

4. Karkischenko N.N., Ryabykh V.P., Karkischenko V.N., Koloskova E.M. Sozdanie gumanizirovannykh myshey dlya farmakotoksikologicheskikh issledovaniy (uspekhi, neudachi I perspektivy) [Creation of humanized mice for pharmacotoxicological research (successes, failures and prospects). Biomedicina [ Journal Biomed]. 2014;1(3):4–22. (In Russian)

5. Karkischenko V.N., Ryabykh V.P., Karkischenko N.N., Dulya M.S., Ezerskiy V.A., Koloskova E.M., Lazarev V.N., Maksimenko S.V., Petrova N.V., Stolyarova V.N., Trubitsyna T.P. Molekulyarno-geneticheskie aspekty tekhnologii polucheniya transgennykh myshej s integrirovannymi genami N-atsetiltransferazy (NAT1 i NAT2) cheloveka [Molecular and genetic aspects of the technology for producing transgenic mice with integrated genes of human N-acetyltransferase (NAT1 and NAT2)]. Biomedicina [ Journal Biomed]. 2016;1:4–17. (In Russian).

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