Cloning and Expression of Brain-Specific Anion Transporter BSAT1 and Isolation of Monoclonal Antibodies to Its Extracellular Fragment

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• 作者：V. P. Baklaushev (12) serpoff@mail.ru
  K. Sh. Kardashova (1)
  O. I. Gurina (1)
  A. V. Leopol’d (2)
  A. V. Semenova (1)
  I. A. Ryabukhin (1)
  V. P. Chekhonin (12)
• 关键词：BSAT1 – monoclonal antibodies – blood ; brain barrier
• 刊名：Bulletin of Experimental Biology and Medicine
• 出版年：2012
• 出版时间：April 2012
• 年：2012
• 卷：152
• 期：6
• 页码：734-738
• 全文大小：584.6 KB
• 参考文献：1. V. P. Ch ekhonin, O. I. Gurina, and T. B. Dmitrieva, Monoclonal Antibodies to Neurospecifi c Proteins [in Russian], Moscow (2007).
  2. F. D. Ca massei, R. Boldrini, A. Jenkner, et al., Pediatr. Dev. Pathol., 6, No. 6, 511-519 (2003).
  3. V. P. Ch ekhonin, O. I. Gurina, T. B. Dmitrieva, et al., Byull. Exp. Biol. Med., 132, No. 2, 772-775 (2001).
  4. R. I. Dm itriev, T. V. Korneenko, A. A. Bessonov, et al., Biochem. Biophys. Res. Commun., 355, No. 4, 1051-1057 (2007).
  5. M. V. Eg orov, A. Tigerstr?m, N. B. Pestov, et al., Protein Expr. Purif., 36, No. 1, 31-39 (2004).
  6. G. K?hler and C. Milstein, Nature, 256, 495-497 (1975).
  7. J. Y. Li, R. J. Boado, and W. M. Pardridge, J. Cereb. Blood Flow Metab., 21, No. 1, 61-68 (2001).
  8. N. B. Pestov and J. Rydstr?m, Nat. Protoc., 2, No. 1, 198-202 (2007).
  9. F. Pizzagal li, B. Hagenbuch, B. Stieger, et al., Mol. Microbiol., 16, No. 10, 2283-2296 (2002).
  10. K. Terpe, Appl. Microbiol. Biotechnol., 60, No. 5, 523-533 (2003).
  11. D. Sugiyama , H. Kusuhara, H. Taniguchi, et al., J. Biol. Chem., 278, No. 44, 43,489-43,495 (2003).
  12. T. Suzuki, T. Onogawa, N. Asano, et al., Mol. Microbiol., 17, No. 7, 1203-1215 (2003).
• 作者单位：1. Department of Fundamental and Applied Neurobiology, V. P. Serbskii State Research Center of Social and Forensic Psychiatry, Moscow, Russia2. Research Center Medical Nanobiotechnologies, Russian State Medical University, Moscow, Russia
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Biomedicine
  Internal Medicine
  Cell Biology
  Pathology
  Laboratory Medicine
  Russian Library of Science
  
• 出版者：Springer New York
• ISSN：1573-8221

文摘

Brain-specifi c anion transporter BSAT1 extracellular fragment (451-557) cDNA was cloned in a vector for prokaryotic expression, a producer E. coli strain was obtained, and recombinant extracellular fragment BSAT1_451-557 was purifi ed and used for immunization of BALB/c mice. Splenic cells from mice with verifi ed immune response were used for hybridoma generation. Several hybridoma clones producing monoclonal antibodies to BSAT1 extracellular fragment were selected. Antibody specifi city was confi rmed by ELISA, immunoblotting with recombinant BSAT1_451-557, and immunofl uorescent BSAT1 assay on rat brain sections and cultured HEK293 cells. It was demonstrated that the obtained antibodies specifi cally bind native rat and human BSAT1 and can be used in both fundamental studies of structures forming the blood-brain barrier and development of targeted transport of diagnostic preparations and drugs across the blood-brain barrier.