We report the development of the first radioimmunoassay for YKL-40, a M(r) = 40 kDa protein which is secreted at high levels by human synovial cells and articular cartilage chondrocytes, and by the human osteosarcoma cell line MG63. This assay uses YKL-40 purified from the conditioned medium of MG63 cells as standard and tracer, and as antigen for immunizing rabbits. With this assay we have discovered high levels of YKL-40 antigen in serum and SF. The molecular weight of serum and SF YKL-40 is identical to purified YKL-40. To evaluate the possible utility of YKL-40 in the assessment of joint disease, we measured YKL-40 in serum and SF of 49 patients with various forms of inflammatory and degenerative joint disease and in the serum of 50 normal adults. The YKL-40 level in serum was significantly higher (P < 0.001) in the patients compared to the normal adults, but there was no difference in serum YKL-40 between the patients with inflammatory joint diseases and OA. The SF levels of YKL-40 were 15-fold higher than serum levels and there was a significant correlation (r = 0.55, P < 0.001) between YKL-40 concentration in SF and serum. Although the tissue distribution of YKL-40 secretion is presently unknown, these observations suggest that a major portion of serum YKL-40 in fact arises from the joint.(ABSTRACT TRUNCATED AT 250 WORDS)