Objective: To purify human VLDL apolipoproteins by middle-pressure liquid chromatography.
Methods: Human VLDLs were isolated by one step density ultracentrifugation. Delipided human VLDL was separated by Sephacryl S-200 molecular sieve chromatography. ApoE was purified by heparin Sepharose CL-6B affinity chromatography. ApoC I ,C II and C III were purified from apoC. fraction by DEAE-Sephacel ion exchange chromatography:
Results: Purified apoE, apoC I, apoC II and apoC III were obtained. SDS-PAGE and immunodiffusion tests indicated the isolated proteins were pure.
Conclusion: We have established a purification procedure for human VLDL apolipoproteins with highly efficiency and simplicity by MPLC.