Establishment of a monoclonal antibody (1/14/16H9) for detection of equine keratan sulfate.

The researchers developed a monoclonal antibody that can accurately detect the presence and measure the concentration of equine keratan sulfate (KS) in blood serum and synovial fluid from horses. The newly developed method could enhance the evaluation of cartilage metabolic activity in horses.

Study Methodology

• The study was conducted using 18 synovial fluid and 48 serum samples collected from both clinically normal horses and those suffering from arthritis.
• The test subjects, BALB/c mice, were immunized with a chondroitinase-ABC-digested proteoglycan monomer derived from equine joint cartilage. Subsequently, monoclonal antibodies were developed with the aid of Sp2/O cells as a fusion partner.
• A unique monoclonal antibody, labelled as MAB1/14/16H9, was identified. This antibody showed high reactivity towards equine KS but did not react with a range of other compounds that include chondroitin sulfate, the core protein of proteoglycan monomers, hyaluronan, heparin, dermatan sulfate, and heparan sulfate.
• The researchers optimized a competitive enzyme-linked immunosorbent assay (ELISA), which deployed the MAB1/14/16H9 antibody to detect and measure KS concentration. The ELISA showed a precision output for the concentration range of 10 to 160 ng/ml. The intra-assay and inter-assay coefficients of variation were 10.0% and 12.7% respectively.

Study Results

• The result of the study showed the successful establishment of MAB 1/14/16H9 as a specific recognition tool for equine KS.
• The optimized ELISA was able to measure the concentration of KS in the synovial fluid and serum samples.
• There was no cross-reactivity observed between the newly established monoclonal antibody and other compounds.

Study Conclusions

• The research resulted in the development of a novel monoclonal antibody and associated method to accurately detect and measure equine KS. This can be applied in blood serum and synovial fluid from horses.
• This development is expected to significantly enhance the evaluation of metabolic activity of cartilage in horses, facilitating a detailed understanding of diseases linked to cartilage health.