Abstract
An antigenic activity in pleural effusions of patients with squamous-cell carcinoma of the lung has been prepared in highly purified form by a 5-step fractionation scheme. The purified substance, designated LuCA (lung cancer antigen), was assessed during the course of the fractionation procedure by a radioimmunometric assay carried out with specific soluble reagents. Sensitive saturation-binding assays showed no or only weak uptake of the 125I-labelled antigen preparation by a panel of antisera specific for known bronchogenic tumour markers, and for normal human serum proteins. The preparation appeared to contain lung-tumour-associated antigens, one of them probably distinct for squamous-cell carcinomas. The antigen fraction consists of acid-soluble glycoproteins, and was demonstrated by SDS-polyacrylamide gel electrophoresis as a single band in the mol. wt region of 43,000. The gel-filtration elution volume appeared to indicate the occurrence of the antigenic activity in multiples of this smaller unit. Pilot radioimmunoassays performed with LuCA and an absorbed specific antiserum suggest the possible suitability of the marker preparation for screening lung-cancer patients.
Highlights
Summary.-An antigenic activity in pleural effusions of patients with squamous-cell carcinoma of the lung has been prepared in highly purified form by a 5-step fractionation scheme
Immunofluorescence, using antisera One of us has described a semi-purified from which antibodies to contaminants glycoprotein with tumour-associated had been removed by absorption
For defining an antiserum titre, 10 1ul (s 8000 ets/min) of a known labelled antigen fraction was pipetted into conical plastic tubes (Sarstedt GmbH, 0 75 ml) which had been rinsed with 0.15% bovine serum albumin (BSA)
Summary
(Louis et al, 1973; Frost et al, 1975; Akeson, 1977). Frost et al (1975) reported an antigen with a mol. wt of 40,000 which was heat and acid stable to a certain extent and showed cationic mobility. For defining an antiserum titre, 10 1ul (s 8000 ets/min) of a known labelled antigen fraction was pipetted into conical plastic tubes (Sarstedt GmbH, 0 75 ml) which had been rinsed with 0.15% bovine serum albumin (BSA). This was followed by the addition of 50 ,ul of the unknown antiserum at 3-fold serial dilutions starting at the concentration 1:20. Day 100 pl goat anti-rabbit antiserum (second antibody) was added, and the tubes were left for another 3 h at 4°C They were centrifuged on an Eppendorf Microfuge at 10,000 rev/min for 5 min in order to separate the bound from the free label.
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