Affinity chromatography with amniotic fluid somatomedin binding protein in the purification of insulin-like growth factor I.
A simplified procedure has been developed for the isolation of insulin-like growth factor I from human plasma by use of affinity chromatography with the somatomedin binding protein. After acidification of human plasma and separation of insulin-like growth factor I and endogenous binding protein by cation exchange chromatography on SP-Sephadex the material was passed through a column packed with pure human amniotic fluid binding protein covalently coupled to Sepharose. The bound insulin-like growth factors I and II were eluted by 1M acetic acid and separated on a Mono S cation exchange column by use of a salt gradient. The 30 micrograms insulin-like growth factor I and 18 micrograms insulin-like growth factor II recovered from 1 liter plasma gave an overall recovery of 30% for insulin-like growth factor I but only 2.5% for insulin-like growth factor II.
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Póvoa
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Wennberg
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Hall
D000596:Amino Acids / Q000032:analysis; D000653:Amniotic Fluid / Q000032:analysis*; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D005260:Female; D006801:Humans; D018969:Insulin-Like Growth Factor Binding Proteins; D011247:Pregnancy; D013002:Somatomedins / Q000302:isolation & purification*
D000596:Amino Acids; D002352:Carrier Proteins; D018969:Insulin-Like Growth Factor Binding Proteins; D013002:Somatomedins