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Characterization of a Human Lung Tumor-associated Antigen and Development of a Radioimmunoassay

Laboratory of Immunodiagnosis, National Cancer Institute, NIH, Bethesda, Maryland 20205

A human lung tumor-associated antigen (LTA), previously isolated from a small cell carcinoma, was further studied after labeling with N-succinimidyl-3-(4-hydroxy-5-[¹²⁵I]iodophenyl)propionate. Two immunoreactive forms of the labeled antigen were observed and isolated after electrophoresis in 7% polyacrylamide gels. These components, referred to as LTA-I and LTA-II in order of mobility, were judged homogeneous by gel electrophoresis, G-200 gel filtration, size exclusion high-per-formance liquid chromatography, and sedimentation velocity analysis. The latter three techniques produced identical profiles for both forms of the LTA. Sephadex chromatography and high-performance liquid chromatography analyses indicated the mass of the antigens to be approximately 140,000 to 150,000 daltons with a D_20,w of 4.2 to 4.3 x 10^-7 sq cm/sec. The s_20,w values for both were 4.5 to 4.6S. Sodium dodecyl sulfate gel electrophoresis of LTA-II gave a single component with a molecular weight of 81,700, while LTA-I had a major component identical in size to LTA-II and two minor components with molecular weights of 50,000 and 27,700, respectively. The isoelectric point of LTA-II (peaks at pH 2.6 and 3.2) generally was more acidic than LTA-I (major component centered at pH 4.7, minor component centered at pH 3.1). A radioimmunoassay, with a useful detection range of from 1 to 100 ng/ml, was developed with LTA-I. This assay was used to determine the concentration of LTA in the sera of normal and lung cancer patients. Fifteen normal sera had a mean of 17 ± 22 (S.D.) ng/ml, with none greater than 83 ng/ml (+3 S.D.). Thirteen lung cancer patients with Stage I (i.e., localized disease) had a mean of 187 ± 219 ng/ml, with the means for 7 of 13 patients being greater than 83 ng/ml; 15 lung cancer patients with Stage III, more extensive disease, had a mean of 277 ± 252 ng/ml, with the means for 12 of 15 patients being greater than 83 ng/ml. This antigen may be useful for the early detection or monitoring of lung cancer.

¹ Present address: Frederick Cancer Research Facility, Bldg. 560, Room 3152, Frederick, Md. 21701. To whom requests for reprints should be addressed.

Received 8/ 7/81. Accepted 11/18/81.