Related Experiment Video
Updated: Sep 6, 2026
Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Identification and analysis of anti-HDL scFv-antibodies obtained from phage display based synthetic antibody library
Priyanka Negi1, Janita Lövgren1, Päivi Malmi1
1Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
Insights
Researchers developed novel antibodies targeting high-density lipoprotein cholesterol (HDL-C) for potential coronary artery disease (CAD) risk assessment. These antibodies show promise as research tools and in diagnostic methods for CAD.
Area of Science:
- Immunology
- Cardiovascular Research
- Biotechnology
Background:
- Plasma high-density lipoprotein cholesterol (HDL-C) levels inversely correlate with atherosclerotic cardiovascular events.
- HDL comprises subpopulations with varying anti-atherogenic properties.
- Coronary artery disease (CAD) diagnosis and risk assessment require improved biomarkers.
Purpose of the Study:
- To isolate coronary artery disease (CAD) specific single-chain variable fragment (scFv) antibodies targeting high-density lipoprotein (HDL).
- To characterize the binding profiles of these novel anti-HDL antibodies.
- To evaluate their potential utility in CAD risk assessment.
Main Methods:
- Phage displayed synthetic antibody libraries were used to enrich for HDL binders from CAD patients.
- Antibodies were affinity purified and characterized using time-resolved fluorescence-based immunoassay.
- Binding to apolipoproteins A-I and A-II, various HDL forms, and plasma HDL was assessed.
Main Results:
- 1200 HDL-binding clones were obtained, yielding 264 unique antibodies after sequencing.
- 61 antibodies were selected for further analysis, revealing diverse binding profiles.
- Several antibodies demonstrated specificity for HDL from CAD patients and recognized HDL in plasma.
Conclusions:
- Novel HDL-recognizing antibodies were successfully isolated using synthetic antibody phage libraries.
- These antibodies exhibit unique binding characteristics, suitable for research applications.
- A subset of these antibodies holds potential for developing diagnostic methods for CAD risk assessment.
Objective:
In epidemiological studies plasma high density lipoprotein cholesterol (HDL-C) levels are found to correlate inversely with atherosclerotic cardiovascular events. HDL consists of different subpopulations and they vary in their anti-atherogenic properties. The aim of this study is to isolate coronary artery disease (CAD) specific anti-HDL scFv-antibodies.
Design And Methods:
To obtain CAD specific HDL binders, we used phage displayed synthetic antibody libraries to enrich specific antibodies against HDL isolated from CAD patients. The antibodies were affinity purified. Their capability to recognize apolipoproteins A-I and A-II, various HDL forms differing in lipid/protein ratios and plasma HDL, was studied using time-resolved fluorescence based immunoassay.
Results:
Using different selection strategies and immunoassay based screening we obtained altogether 1200 clones displaying HDL binding activity. By sequencing 337, we identified 264 unique antibodies against HDL. A set of 61 antibodies were selected for further analysis. We found a variety of antibodies with different binding profiles, including apoA-I binding antibodies either in lipid-dependent or lipid-independent manner and binders against apoA-II. Several antibodies were able to discriminate between HDL derived from CAD patients and healthy controls. A majority of the antibodies were immunoreactive with HDL in plasma.
Conclusion:
The novel HDL recognizing antibodies isolated from synthetic antibody phage library have displayed interesting HDL-binding characteristics suggesting that, in addition to use as research tools, a part of them might be useful for the development of diagnostic methods for CAD risk assessment.
More Related Videos
12:31Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity
Published on: May 1, 2018
10:17Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
Published on: January 14, 2020
