Phage-displayed recombinant single-chain antibody fragments with high affinity for cholesteryl ester transfer protein (CETP): cDNA cloning, characterization and CETP quantification
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Andreas Ritsch
Abstract
Cholesteryl ester transfer protein (CETP) greatly affects the metabolism of all lipoprotein classes including low-density lipoprotein (LDL) and high-density lipoprotein (HDL), bothknown 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 KD 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 withh igh 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.
Copyright © 2004 by Walter de Gruyter GmbH & Co. KG
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- Meetings and Awards
Articles in the same Issue
- Phage-displayed recombinant single-chain antibody fragments with high affinity for cholesteryl ester transfer protein (CETP): cDNA cloning, characterization and CETP quantification
- Stem cell factor and macrophage-colony stimulating factor in patients with pancreatic cancer
- Modulation of plasma fibrinogen levels in acute-phase response after hepatectomy
- Anti-nucleosome, anti-chromatin, anti-dsDNA and anti-histone antibody reactivity in systemic lupus erythematosus
- Expression of activation molecules in neutrophils, monocytes and lymphocytes from patients with unstable angina treated with stent implantation
- Acid ribonuclease and alkaline ribonuclease isoenzymes in plasma of patients with decreased glomerular filtration rate
- Is calcium oxalate nucleation in postprandial urine of males with idiopathic recurrent calcium urolithiasis related to calcium phosphate nucleation and the intensity of stone formation? Studies allowing insight into a possible role of urinary free citrate and protein
- Significance of consecutive international normalized ratio (INR) outcomes using statistical control rules in long-term anticoagulated patients. Optimization of laboratory monitoring and interpretation of borderline measurements
- Clinical trials and p-values, beware of the extremes
- Age dependence of serum β-N-acetylhexosaminidase (NAG) activity
- Elevated plasma total homocysteine in centenarians
- New methodology of influential point detection in regression model building for the prediction of metabolic clearance rate of glucose
- Intra-laboratory analytical variability of biochemical markers of fibrosis (Fibrotest) and activity (Actitest) and reference ranges in healthy blood donors
- The glutathione S-transferase polymorphisms in a control population and in Alzheimer's disease patients
- Near-patient testing for infection using urinalysis and immuno-chromatography strips
- Long-distance mountain biking does not disturb the measurement of total, free or complexed prostate-specific antigen in healthy men
- Development of a new colorimetric method for protein S activity measurement
- Re.: Correction to the IFCC primary reference method for the measurement of catalytic activity concentration of enzymes at 37C- part 2: reference procedure for the measurement of creatine kinase. Clin Chem Lab Med 2002;40:635-42
- Meetings and Awards
