Isolation and characterization of anti ROR1 single chain fragment variable antibodies using phage display technique.

Publisher: IOS Press Country of Publication: Netherlands NLM ID: 9711270 Publication Model: Print Cited Medium: Internet ISSN: 1875-869X (Electronic) Linking ISSN: 10932607 NLM ISO Abbreviation: Hum Antibodies Subsets: MEDLINE

Publication: Amsterdam : IOS Press
Original Publication: Wilton, CT : Forefront Pub., c1997-

Cell Surface Display Techniques* ; Peptide Library*; Antibodies, Monoclonal/*isolation & purification ; Receptor Tyrosine Kinase-like Orphan Receptors/*immunology ; Single-Chain Antibodies/*isolation & purification; Antibodies, Monoclonal/chemistry ; Antibodies, Monoclonal/genetics ; Antibody Specificity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Gene Expression ; Humans ; Protein Binding ; Protein Domains ; Receptor Tyrosine Kinase-like Orphan Receptors/chemistry ; Receptor Tyrosine Kinase-like Orphan Receptors/genetics ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/immunology ; Single-Chain Antibodies/chemistry ; Single-Chain Antibodies/genetics

Receptor tyrosine kinase-like orphan receptor (ROR1) belongs to one of the families of receptor tyrosine kinases (RTKs). RTKs are involved in the various physiologic cellular functions including proliferation, migration, survival, signaling and differentiation. Several RTKs are deregulated in various cancers implying the targeting potential of these molecules in cancer therapy. ROR1 has recently been shown to be expressed in various types of cancer cells but not in normal adult cells. Hence a molecular inhibitor of extracellular domain of ROR1 that inhibits ROR1-cell surface interaction is of great therapeutic importance. In an attempt to develop molecular inhibitors of ROR1, we screened single chain variable fragment (scFv) phage display libraries, Tomlinson I + J, against one specific synthetic oligopeptide from extracellular domain of ROR1 and selected scFvs were characterized using various immunological techniques. Several ROR1 specific scFvs were selected following five rounds of panning procedure. The scFvs showed specific binding to ROR1 using immunological techniques. Our results demonstrate successful isolation and characterization of specific ROR1 scFvs that may have great therapeutic potential in cancer immunotherapy.

Keywords: ROR1; phage display technique; receptor tyrosine kinases; scFv

0 (Antibodies, Monoclonal)
0 (Peptide Library)
0 (Recombinant Proteins)
0 (Single-Chain Antibodies)
EC 2.7.10.1 (ROR1 protein, human)
EC 2.7.10.1 (Receptor Tyrosine Kinase-like Orphan Receptors)

Date Created: 20170128 Date Completed: 20170310 Latest Revision: 20170310

20250114

10.3233/HAB-170310

28128766