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Detergent solubilization and partial purification of tumor specific surface and transplantation antigens from
Abstract:
A solubilization technique employing 0.5% Triton X-100 was developed to obtain both SV40 virus (SV40)-induced tumor-specific surface antigen(s) (TSSA) from SV40-transformed mouse cells, as determined by a serum-mediated microcytolytic assay, and tumor-specific transplantation antigen(s) (TSTA), as determined by in invivo experiments. High yields (approximately 50%) of TSSA were obtained in whole-cell extracts and also after ammonium sulfate fractionation. Additional fractionation of a 30-50% ammonium sulfate fraction by gel exclusion chromatography on Sephadex G-150 resulted in two pooled fractions which contained TSSA activity. The first eluted close to the void volume, and the second in the 45,000 molecular weight region. The various TSSA active fractions were also active in vivo TSTA tests. Detergent solubilization provides a suitable technique to recover the SV40-induced antigens in good yield, and apparently in intact form.
Insights
Researchers developed a method using Triton X-100 to extract simian virus 40 (SV40)-induced tumor antigens. This technique successfully isolated both surface (TSSA) and transplantation (TSTA) antigens from SV40-transformed cells in high yields.
Area of Science:
- Immunology
- Virology
- Biochemistry
Background:
- SV40-induced tumors express unique antigens.
- Efficient extraction of these antigens is crucial for immunological studies.
Purpose of the Study:
- To develop a solubilization technique for isolating SV40-induced tumor-specific antigens (TSSA and TSTA).
- To characterize the properties and yield of extracted antigens.
Main Methods:
- Solubilization of SV40-transformed mouse cells using 0.5% Triton X-100.
- Serum-mediated microcytolytic assays for TSSA detection.
- In vivo experiments for TSTA determination.
- Ammonium sulfate fractionation and Sephadex G-150 gel exclusion chromatography.
Main Results:
- High yields (approx. 50%) of TSSA were obtained from whole-cell extracts and fractionated samples.
- Two active fractions containing TSSA were identified by chromatography, eluting near the void volume and at a 45,000 MW region.
- Fractions active for TSSA also demonstrated TSTA activity in vivo.
Conclusions:
- Detergent solubilization with Triton X-100 is an effective method for recovering SV40-induced antigens.
- The technique yields antigens in apparently intact form and in good quantity.
- This method facilitates further study of SV40 tumor antigens.