title: Development of glycan specific lectin based immunoassay for detection of prostate specific antigen.
abstract: We describe an analytical approach for the detection and verification of glycosylation patterns of prostate specific antigen (PSA), a key biomarker currently used for understanding the onset and prognosis of prostate cancer. PSA has been purified from the human seminal plasma and total PSA from prostate cancer sera. PSA is a monomeric glycoprotein with an apparent molecular mass 28040.467 Da, which exhibits a characteristic protease activity against casein and gelatin. Its optimal protease activity is centered on neutral pH. Peptide mass fingerprint analysis of the purified PSA has yielded peptides that partially match with known database sequences (Uniprot ID P07288). Tryptic digestion profile of isolated PSA, infer the exclusive nature of PSA and may be additive molecule in the dictionary of seminal proteins. Surface plasmon resonance and lectin immunoassay revealed direct interaction between a newly developed anti-PSA monoclonal antibody (C4E6) and PSA. A lectin based immunoassay is reported here which was achieved with the C4E6 anti-PSA antibody and biotinylated plant lectins. This investigation provides an alternative method to isolate and quantify PSA with altered glycosylation which might be seen in the prostate cancer and developing a lectin based immunoassay to detect PSA in serum of prostate cancer patients.
authors: Laboratory of Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, India; Department of Medical Biotechnology, MGM Institute of Health Sciences, Kamothe, Navi Mumbai 410209, India.
authors: Paresh B
authors: PB
authors: Bhanushali
authors: Laboratory of Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, India; Department of Biochemistry, National Institute for Research in Reproductive Health, Jehangir Merwanji Street, Parel, Mumbai 400012, India. Electronic address: sham83badgujar@gmail.com.
authors: Shamkant B
authors: SB
authors: Badgujar
authors: Laboratory of Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, India.
authors: Mukesh M
authors: MM
authors: Tripathi
authors: Laboratory of Native Proteins, Research and Development Division, Yashraj Biotechnology Ltd., Navi Mumbai 400705, India.
authors: Sanjeev
authors: S
authors: Gupta
authors: Department of Radiation Oncology, Tata Memorial Centre, Mumbai 400012, India.
authors: Vedang
authors: V
authors: Murthy
authors: Laboratory of Membrane Biology, National Centre for Cell Science, Savitribai Phule Pune University Campus, Pune 411007, India.
authors: Musti V
authors: MV
authors: Krishnasastry
authors: Department of Medical Biotechnology, MGM Institute of Health Sciences, Kamothe, Navi Mumbai 410209, India.
authors: Chander P
authors: CP
authors: Puri
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D037102:Lectins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D017430:Prostate-Specific Antigen / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000276:immunology; D012661:Semen / Q000378:metabolism; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D037102:Lectins; D011134:Polysaccharides; D017430:Prostate-Specific Antigen
keywords: Biomarkers; Glycosylation; Lectin-based immunoassay; Prostate cancer; Prostate specific antigen