Avtor/Urednik		Bojanič, Vesna
Naslov		Priprava monoklonskih protiteles proti humanemu TNF alfa, primernih za uporabo v medicinski diagnostiki
Tip		monografija
Kraj izdaje		Ljubljana
Založnik		Medicinska fakulteta
Leto izdaje		1998
Obseg		str. 67
Jezik		slo
Abstrakt		TNFalpha is a pleiotropic cytokine primarily produced by activated macrophages. The activity of TNFalpha includes cytotoxicity, immunoregulatorry activity, mitogenic activity and transcriptional regulation of many genes. TNFalpha exists as a homotrimer of 17 kDa subunit polypeptides, and the formation of a trimer structure is likely to be essential for its biological activity. The activity of TNFalpha is mediated through two distinct receptors, one with a molecular mass of 55 kDa (TNF-RI or p55) and another with a molecular mass of 75 kDa (TNF-RII or p 75). Most cell types and tissues express one or both of two receptor types. There is some evidence for clinical importance of TNFalpha and both soluble receptors, thus we produced monoclonal antibodies and developed our own ELISA for quantitative determination of human TNFalpha for the purposes of medical diagnostics, and ELISA for study the structure of the molecule as well as for monitoring the biotechnological expression of human recombinant TNFalpha analogues. To obtain different types of monoclonal antibodies we immunized BALB/c mice with native recombinant human TNFalpha analogue, Lk800, termostable recombinant human TNFalpha, Lk811 and complexes of a soluble receptor p55 or p75 with termostable less toxic mutants of TNFalpha, Lk802 or Lk811. Mice spleen cells isolated from immunized animals were fused P3-NS1/1-Ag4-1 mouse myeloma cell line. Supernatants were screened for specific antibody production production by ELISA. We selected the best producers and purified monoclonal antibodies by affinity chromatography on protein G-Sepharose. We compared monoclonal antibodies with polyclonal ones which were produced in rabbits. A pair of monoclonal antibodies, F7/2 and C9/8, which recognize different epitopes on the native TNFalpha molecule were used to develop a sensitive sandwich ELISA which might be used in medical diagnostics. (Abstracts truncated at 2000 characters).
Deskriptorji		TUMOR NECROSIS FACTOR ANTIBODIES, MONOCLONAL MELANOMA MICE, INBRED BALB C CELL LINE, TRANSFORMED ENZYME-LINKED IMMUNOSORBENT ASSAY CHROMATOGRAPHY, AFFINITY HYBRID CELLS ELECTROPHORESIS ISOELECTRIC FOCUSING BLOTTING, WESTERN NEUTRALIZATION TESTS