Polyreactivity of Monoclonal Antibodies Produced Against Thyroid Stimulating Hormone (hTSH)

Abstract

Monoclonal antibodies against human thyroid stimulating hormone (hTSH) are among the key reagents required in the development of an immunoradiometric assay (IRMA) for hTSH in serum. In this study we have produced and characterized twelve hTSH monoclonal antibodies. Hybridomas were generated by fusion of B-lymphocytes from mice immunized with hTSH and myeloma cells. Clones producing antibodies against hTSH were selected using ELISA (Enzyme Linked Immunosorbent Assay). Antibodies from the selected and cloned hybridoma cells were purified by affinity chromatography and their reactivities were tested by ELISA against a panel of antigens, i.e., hTSH, bovine serum albumin (BSA), and milk protein, and also by studying the binding of these monoclonal antibodies with the radioiodinated antigens (hTSH, BSA, and milk protein). It was observed that some of the hTSH monoclonal antibodies produced were polyreactive, reacting with hTSH as well as with unrelated antigen BSA, while others were monoreactive, reacting only to hTSH.

Highly Sensitive Chemiluminescent Analysis of Residual Bovine Serum Albumin (BSA) Based on a Pair of Specific Monoclonal Antibodies and Peroxyoxalate–glyoxaline–PHPPA Dimer Chemiluminescent System in Vaccines

“…BSA has numerous biochemical applications, including protein microarray technology [3], lysate microarray technology [4], enzyme-linked immunosorbent assay (ELISA) [5,6], immunoblots as a blocking agent during western blotting to reduce background and non-specific binding [7], and immunohistochemistry [8]. Moreover, it is generally used in vaccine delivery studies as a model protein [9][10][11][12][13][14], owing to its ease of assay, well-known immunogenicity pattern, well-known physicochemical properties, low cost, and general availability.…”

BSA/HSA ratio modulates the properties of Ca2+-induced cold gelation scaffolds

“…BSA is an acidic single polypeptide protein extensively used in pharmaceutical industry for a variety of applications. It is used as a stabilizer [6], a blocking agent in various immunoassays [7], in microarray technologies [8], in cell culture medium as growth supplement for viral vaccine manufacturing [9] and in nanoparticles for drug delivery [10]. Despite widely used there are studies showing BSA to be a trigger for allergic reactions [11,12].…”

B-1 Cell-Derived Monoclonal Antibodies and Costimulatory Molecules

“…Some monoclonal Abs had the capacity to bind to a variety of different Ags [12,13]. Notkins [10] found that 10% to 15% of the cells made broadly reactive Abs, which are now referred to as polyreactive Abs (natural Abs); in addition, the majority of polyreactive Abs are of the IgM isotype.…”