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001-es BibID:	BIBFORM069265
Első szerző:	Wang, Dongdong
Cím:	Antigen Identification and Characterization of Lung Cancer Specific Monoclonal Antibodies Produced by mAb Proteomics / Wang Dongdong, Hincapie Marina, Guergova-Kuras Mariana, Kadas Janos, Takacs Laszlo, Karger Barry L.
Dátum:	2010
ISSN:	1535-3893
Megjegyzések:	A mass spectrometric (MS)-based strategy for antigen (Ag) identification and characterization of globallyproduced monoclonal antibodies (mAbs) is described. Mice were immunized with a mixture of nativeglycoproteins, isolated from the pooled plasma of patients with nonsmall cell lung cancer (NSCLC), togenerate a library of IgG-secreting hybridomas. Prior to immunization, the pooled NSCLC plasma wassubjected to 3-sequential steps of affinity fractionation, including high abundant plasma proteindepletion, glycoprotein enrichment, and polyclonal antibody affinity chromatography normalization.In this paper, to demonstrate the high quality of the globally produced mAbs, we selected 3 mAbs ofhigh differentiating power against a matched, pooled normal plasma sample. After production of largequantities of the mAbs from ascites fluids, Ag identification was achieved by immunoaffinity purification,SDS-PAGE, Western blotting, and MS analysis of in-gel digest products. One antigen was found to becomplement factor H, and the other two were mapped to different subunits of haptoglobin (Hpt). The2 Hpt mAbs were characterized in detail to assess the quality of the mAbs produced by the globalstrategy. The affinity of one of the mAbs to the Hpt native tetramer form was found to have a KD ofroughly 10-9 M and to be 2 orders of magnitude lower than the reduced form, demonstrating thepower of the mAb proteomics technology in generating mAbs to the natural form of the proteinsin blood. The binding of this mAb to the -chain of haptoglobin was also dependent on glycosylationon this chain. The characterization of mAbs in this work reveals that the global mAb proteomicsprocess can generate high-quality lung cancer specific mAbs capable of recognizing proteins intheir native state.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban monoclonal antibody mAb proteomics antibody characterization glycoprotein haptoglobin conformational epitope lung cancer
Megjelenés:	Journal Of Proteome Research. - 9 : 4 (2010), p. 1834-1842. -
További szerzők:	Hincapie, Marina Guergova-Kuras, Mariana Kádas János (1976-) (molekuláris biológus, biokémikus, kertészmérnök) Takács László (1955-) Karger, Barry
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