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- Title
Phage-displayed recombinant single-chain antibody fragments with high affinity for cholesteryl ester transfer protein (CETP): cDNA cloning, characterization and CETP quantification.
- Authors
Ritsch, Andreas; Ebenbichler, Christoph; Naschberger, Elisabeth; Schgoer, Wilfried; Stanzl, Ursula; Dietrich, Hermann; Heinrich, Peter C.; Saito, Kazunori; Patsch, Josef R.
- Abstract
Cholesteryl ester transfer protein (CETP) greatly affects the metabolism of all lipoprotein classes including low-density lipoprotein (LDL) and high-density lipoprotein (HDL), both known to constitute powerful risk factors for coronary artery disease (CAD). We now report the successful first cloning and characterization of single-chain antibody fragments specific for CETP. A recombinant phage display library was generated using spleen mRNA isolated from BALB/c mice that had been immunized with highly purified CETP. Screening of the library yielded two single-chain antibody fragments with high affinity for CETP, termed 1CL8 and 1CL10, displaying respective K D values of 4.36×10-9 M and 4.64×10-9 M as determined by affinity sensor technology. Amino acid sequence comparison indicated the complementarity determining regions of the respective heavy chains to be responsible for CETP high affinity binding. Fragment 1CL8 was successfully employed in clinical chemical quantification systems that uncovered an association in humans between plasma CETP concentration and total body fat mass (r=0.50, p<0.002). Because of the demonstrated superb CETP capturing capacity, combined with high binding affinity to CETP, ready access and unlimited supply, 1CL8 and 1CL10 are expected to prove powerful tools for studies on the role of CETP in atherogenesis.
- Subjects
CLONING; RECOMBINANT antibodies; ESTERS; ENZYME-linked immunosorbent assay; HIGH density lipoproteins; LOW density lipoproteins; AFFINITY chromatography; MESSENGER RNA
- Publication
Clinical Chemistry & Laboratory Medicine, 2004, Vol 42, Issue 3, p247
- ISSN
1434-6621
- Publication type
Article
- DOI
10.1515/CCLM.2004.046