rise and fall of an anti-muc1 specific antibody的兴衰anti-muc1特异性抗体.pdfVIP

Rise and Fall of an Anti-MUC1 Specific Antibody Holger Thie1.¤a, Lars Toleikis1.¤b, Jiandong Li1¤c, Reinhard von Wasielewski2¤d, Gunther Bastert3¤e, 1 4 4 ´ ´ 4 Thomas Schirrmann , Isabel Tourais Esteves , Christian K. Behrens , Benedict Fournes , Nathalie 4 4 1 ¨ 1 Fournier , Christophe de Romeuf , Michael Hust , Stefan Dubel * ¨ ¨ 1Technische Universitat Braunschweig, Institut fur Biochemie und Biotechnologie, Braunschweig, Germany, 2 Hannover Medical School, Institute of Pathology, Hannover, ¨ Germany, 3 Universitatsfrauenklinik Heidelberg, Heidelberg, Germany, 4 LFB Biotechnologies - Research Department, Lille, France Abstract Background: So far, human antibodies with good affinity and specificity for MUC1, a transmembrane protein overexpressed on breast cancers and ovarian carcinomas, and thus a promising target for therapy, were very difficult to generate. Results: A human scFv antibody was isolated from an immune library derived from breast cancer patients immunised with MUC1. The anti-MUC1 scFv reacted with tumour cells in more than 80% of 228 tissue sections of mamma carcinoma samples, while showing very low reactivity with a large panel of non-tumour tissues. By mutagenesis and phage display, affinity of scFvs was increased up to 500fold to 5,7 610210 M. Half-life in serum was improved from below 1 day to more than 4 weeks and was correlated with the dimerisation tendency of the individual scFvs. The scFv bound to T47D and MCF-7 mammalian cancer cell lines were recloned into the scFv-Fc and IgG format resulting in decrease of affinity of one bind