Novel Strategy for Selection of Monoclonal Antibodies Against Highly Conserved Antigens: Phage Library Panning Against Ephrin-B2 Displayed on Yeast

Autor: Yogindra Vedvyas, Xuebo Hu, Moonsoo M. Jin, Chang-Il Hwang, Xiaoling Gu, Tanwi Kaushik, Alexander Yu. Nikitin, Xiaoyue Chen
Rok vydání: 2012
Předmět:
lcsh:Medicine
Biochemistry
Conserved sequence
Mice
Engineering
0302 clinical medicine
Cricetinae
Yeasts
lcsh:Science
Internalization
Cells
Cultured
Conserved Sequence
media_common
0303 health sciences
Multidisciplinary
biology
Antibodies
Monoclonal
3. Good health
030220 oncology & carcinogenesis
embryonic structures
Medicine
Antibody
Research Article
Biotechnology
Protein Binding
Histology
animal structures
Science Policy
medicine.drug_class
Immunoprecipitation
media_common.quotation_subject
Immunology
Molecular Sequence Data
Receptor
EphB4
Mice
Nude
Bioengineering
Ephrin-B2
CHO Cells
Monoclonal antibody
Models
Biological
03 medical and health sciences
Cricetulus
Antigen
Peptide Library
medicine
Animals
Humans
Amino Acid Sequence
Antigens
Panning (camera)
Peptide library
Biology
030304 developmental biology
Sequence Homology
Amino Acid
lcsh:R
Proteins
HCT116 Cells
Xenograft Model Antitumor Assays
Molecular biology
biological factors
High-Throughput Screening Assays
Technology Development
HEK293 Cells
nervous system
biology.protein
Clinical Immunology
lcsh:Q
sense organs
Zdroj: PLoS ONE, Vol 7, Iss 1, p e30680 (2012)
PLoS ONE
ISSN: 1932-6203
Popis: Ephrin-B2 is predominately expressed in endothelium of arterial origin, involved in developmental angiogenesis and neovasculature formation through its interaction with EphB4. Despite its importance in physiology and pathological conditions, it has been challenging to produce monoclonal antibodies against ephrin-B2 due to its high conservation in sequence throughout human and rodents. Using a novel approach for antibody selection by panning a phage library of human antibody against antigens displayed in yeast, we have isolated high affinity antibodies against ephrin-B2. The function of one high affinity binder (named as 'EC8') was manifested in its ability to inhibit ephrin-B2 interaction with EphB4, to cross-react with murine ephrin-B2, and to induce internalization into ephrin-B2 expressing cells. EC8 was also compatible with immunoprecipitation and detection of ephrin-B2 expression in the tissue after standard chemical fixation procedure. Consistent with previous reports on ephrin-B2 induction in some epithelial tumors and tumor-associated vasculatures, EC8 specifically detected ephrin-B2 in tumors as well as the vasculature within and outside of the tumors. We envision that monoclonal antibody developed in this study may be used as a reagent to probe ephrin-B2 distribution in normal as well as in pathological conditions and to antagonize ephrin-B2 interaction with EphB4 for basic science and therapeutic applications.
Databáze: OpenAIRE
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