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Development of a monoclonal antibody against strains of human nanoparticles
Author(s) -
Kraemer Maria A.,
Kumar Vivek,
FarellBaril Gerard,
Miller Virginia M.,
Beito Thomas G.,
Cockerill Franklin R.,
Lieske John C.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a577-a
Subject(s) - monoclonal antibody , western blot , antibody , microbiology and biotechnology , lysis , antigen , kidney , monoclonal , chemistry , biology , immunology , biochemistry , endocrinology , gene
Replicating, self‐calcifying nano‐sized particles have been isolated and cultured by our group and others from human kidney stones and arterial calcifications, suggesting that these particles may contribute to pathological calcification. As no antibodies are available against nanoparticles (NP) isolated from humans, and in order to facilitate further study, we sought to develop monoclonal antibodies against our strains. NP’s were isolated and cultured from human kidney stones and calcified arterial tissue collected as surgical waste, using published techniques. Lysate prepared from decalcified and sonicated NP’s were injected into mice (n = 5/strain). After 28 days, blood was collected and screened for the presence of antibodies against each strain of lysate using ELISA and Western blot. Splenocytes from mice with high antibody titres were fused with the F/O myeloma cell line and then screened by ELISA against NP lysate; reactive clones were then screened via Western blot. Media used to grow the NP’s served as the negative control antigen. Numerous clones reacted against NP‐lysate of the kidney stone strain by ELISA. By Western blot, this monoclonal antibody recognized a 120 kDa protein that was not detected in the control antigen, serum‐containing NP‐media. Therefore, human renal stone NP‐lysates invoke an immunological response in mice. This resulting monoclonal antibody could be used to develop diagnostic tests for the presence of these particles in human blood, urine, and tissue.