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Camel heavy chain antibodies against prostate-specific membrane antigen.
- Source :
-
Hybridoma (2005) [Hybridoma (Larchmt)] 2012 Dec; Vol. 31 (6), pp. 424-9. - Publication Year :
- 2012
-
Abstract
- Prostate-specific membrane antigen (PSMA), a type II integral membrane glycoprotein, is highly overexpressed in all forms of prostate cancer tissues. It has also been demonstrated in a wide range of neovasculature of non-prostatic solid tumors, including bladder, pancreas, lung, kidney, colorectal, and gastric cancers. Given the unique expression of PSMA, it is considered an alluring target for antibody-based imaging and therapy of cancer. In the present study, the production and characterization of camel heavy chain antibodies (HCAbs) specific for the external domain of the PSMA are reported. Due to the absence of the CH1 domain, HCAbs are smaller than their counterparts in conventional antibodies. In this study, camel antibodies were generated through immunization of Camelus dromedarius with a synthetic 28 amino acid peptide corresponding to the external surface domain of antigen and PSMA-expressing cell lines. Different binding properties to protein A and protein G affinity columns were deployed to separate three subclasses of camel IgG. The affinity purified HCAbs bound selectively to the synthetic peptide in enzyme linked immunosorbent assay (ELISA) and reacted specifically with PSMA-expressing cell line through immunocytochemistry study. Currently, we are attempting to develop recombinant variable domain of these heavy chain antibodies (VHH or nanobody) for tumor imaging and cancer therapy.
- Subjects :
- Animals
Antibodies isolation & purification
Antibody Specificity
Antigens, Surface metabolism
Camelus
Cell Line, Tumor
Chromatography, Affinity
Enzyme-Linked Immunosorbent Assay
Glutamate Carboxypeptidase II metabolism
Humans
Immunoglobulin Heavy Chains biosynthesis
Immunoglobulin Heavy Chains isolation & purification
Male
Peptide Fragments chemistry
Peptide Fragments immunology
Prostatic Neoplasms
Protein Binding
Serum Albumin, Bovine chemistry
Antibodies chemistry
Antigens, Surface immunology
Glutamate Carboxypeptidase II immunology
Immunoglobulin Heavy Chains chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1557-8348
- Volume :
- 31
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Hybridoma (2005)
- Publication Type :
- Academic Journal
- Accession number :
- 23244321
- Full Text :
- https://doi.org/10.1089/hyb.2012.0048