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A rapid, large scale purification procedure for gibbon interleukin 2.
Journal of Immunology (Baltimore, Md. : 1950)
|August 1, 1983
Summary
This study successfully purified Interleukin 2 (IL 2) from a gibbon T cell line, yielding highly active IL 2 for further research. The efficient purification process ensures high recovery and specific activity, aiding biochemical and immunologic studies.
Area of Science:
- Immunology
- Biochemistry
- Cell Biology
Background:
- Interleukin 2 (IL 2) is a crucial lymphokine for T cell proliferation.
- Constitutive IL 2 production by cell lines simplifies purification efforts.
- Previous methods for IL 2 purification were less efficient.
Purpose of the Study:
- To develop a rapid and efficient method for purifying large quantities of active Interleukin 2 (IL 2).
- To characterize the molecular properties of purified IL 2.
- To assess the biological activity of purified IL 2.
Main Methods:
- Purification of IL 2 from MLA144 gibbon T cell line conditioned medium.
- Utilized trimethylsilyl-controlled pore glass batch purification and reversed-phase high-pressure liquid chromatography.
- Analyzed purified IL 2 using two-dimensional isoelectric focusing-SDS-polyacrylamide gel electrophoresis.
Main Results:
- Achieved 70-100% recovery of IL 2 activity, yielding ≥ 2 X 10(6) units per 15 liters.
- Purified IL 2 exhibited high specific activity (0.5-1 X 10(8) U/mg protein).
- Identified four active molecular forms of IL 2 (three at 16 kDa, one at 15 kDa) with varying isoelectric points.
Conclusions:
- A robust purification protocol for Interleukin 2 (IL 2) was established.
- The purified IL 2 demonstrated potent biological activity, supporting lymphocyte growth.
- This efficient purification method facilitates advanced biochemical and immunological investigations of IL 2.