Large scale purification of intralumenal alkaline phosphatase from calf intestine by immunosorbent affinity chromatography
Schwenzer, B.; Neef, B.; Bublitz, R.; Ehle, H.; Horn, A.
Biomedica Biochimica Acta 46(1): 15-21
1987
ISSN/ISBN: 0232-766X PMID: 3107547 Document Number: 291211
Electrophoretically and immunologically homogeneous alkaline phosphatase (AP) can be obtained by a simple and rapid two-step procedure. Step 1: Immunosorbent affinity chromatography using immobilized polyclonal anti-AP-antibodies and elution of the bound enzyme by triethylamine solution, pH 11.4. After affinity chromatography the specific activity of the extracted crude material increases from 3.5 to 793 IU/mg. Step 2: Final purification to a specific activity of 1524 IU/mg by DEAE-cellulose ion exchange chromatography. One investigator is able to purify 100 mg AP within 3 days. The overall recovery is 65%. Methods for characterization and selection of anti-AP-antiserum using immunoinhibition of AP are described.