Studies on the isolation and properties of human and hog intrinsic factor

Steele, James (1976) Studies on the isolation and properties of human and hog intrinsic factor. PhD thesis, University of Glasgow.

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Abstract

The main aim of this research work was to isolate pure preparations of both human and hog Intrinsic Factor and examine some of their physico-chemical and immunological properties. The method chosen to isolate pure I.F. was affinity chromatography, in particular the method used by Allen and co-workers in America. Their method was studied and modified, mainly by the use of lower molarity guanidine-HC1 to elute the affinity chromatography column, thus providing a method of separating I.F. from non-I.F. binders. The results gained in the present work by amino acid analysis and gel filtration were similar to those gained by Allen except that the carbohydrate composition of the hog I.F. did not contain any galactosomine and there was no appearance of any dimerised or oligomerised form of I.F. The molecular weight for both human and hog I.F. was approximately 60,000 and the other physico-chemical properties were similar for both preparations. The two were not identical immunologically, since the Type II antibody, which was raised to human I.F, would not react with hog I.F. Both human and hog I.F. preparations were successfully labelled with [125I] using the Chloramine T method and this preparation was used to compare the properties of I.F. with and without the addition of B12. Methods for possible elucidation of I.F. mediated B12 absorption from the intestine are suggested using a peroxidase linked to specific I.F. antibody staining technique and also a specific radioimmunoassay for very small quantities of I.F. A routine clinical radioassay for B12 is also described.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Additional Information: Adviser: J F Adams
Keywords: Immunology
Date of Award: 1976
Depositing User: Enlighten Team
Unique ID: glathesis:1976-72953
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 11 Jun 2019 11:06
Last Modified: 11 Jun 2019 11:06
URI: http://theses.gla.ac.uk/id/eprint/72953

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