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dc.identifier.citationAlmåsbak, Hilde. A single-chain-Fv-based immunofluorometric assay specific for the CEA variant NCA-2. Hovedoppgave, University of Oslo, 2004en_US
dc.description.abstractThe disagreement between various immunoassays for the tumour marker carcinoembryonic antigen (CEA) is well known, and the discrepancies are explained by the co-measurement of the structurally related non-specific cross-reacting antigen 2 (NCA-2). Until now, this hypothesis has only been proven indirectly, as no NCA-2 specific assay has been available. This thesis describes the production of a high affinity single-chain antibody fragment (scFv) raised by using phage display technology. The scFv was used to construct an immunometric assay that specifically recognises NCA-2. A phagemid library was constructed from splenic mRNA obtained from a mouse immunised with NCA-2 purified from human meconium. Following phage rescue and three rounds of selection on solid phase coated with NCA-2, several clones, which displayed scFvs with high specificity and affinity for NCA-2, were isolated. These were sub-cloned into an expression vector for high-level expression of soluble scFv. Based on BIAcore analysis, a scFv with low koff and a KD of 10-10 mol/l was selected for europium labelling and employed as the tracer molecule for the NCA-2 - specific immunofluorometric assay. This novel assay was applied for the quantification of NCA-2 in sera from 35 patients with elevated CEA levels. The measured NCA-2 correlated fairly well (R2=0.82) with the values obtained by indirect methods based on the differences between serum values measured by NCA-2 cross-reactive and CEA specific immunoassays.nor
dc.titleA single-chain-Fv-based immunofluorometric assay specific for the CEA variant NCA-2en_US
dc.typeMaster thesisen_US
dc.creator.authorAlmåsbak, Hildeen_US
dc.identifier.bibliographiccitationinfo:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft.au=Almåsbak, Hilde&rft.title=A single-chain-Fv-based immunofluorometric assay specific for the CEA variant NCA-2&rft.inst=University of Oslo&rft.date=2004&rft.degree=Hovedoppgaveen_US
dc.contributor.supervisorElisabeth Pausen_US

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