Method for the immunological determination of heparan sulfate-proteoglycan and process for the preparation and purification of heparan sulfate-proteoglycan suitable for this purpose from tissues containing basal membrane
AssigneeHoechst Aktiengesellschaft
InventorsRupert Timpl, Mats Paulsson, Dietrich Brocks
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Abstract
The invention relates to a method for the immunological determination of basal membrane low-density heparan sulfate-proteoglycan in body fluids, and to the preparation or obtaining of a low-density heparan sulfate-proteoglycan suitable for this purpose, and of the corresponding highly specific antibodies.
Claims12
A process for the preparation of substantially pure low-density (.delta.=1.33-1.39) heparan sulfate-proteoglycan of about 620-680 kd, which comprises:
- a) extraction of basal membrane tissue from human placenta in the presence of protease inhibitors,
- b) extraction of said low-density heparan sulfate-proteoglycan from said extracted tissue into an aqueous phase with chaotropic salt, and
- c) purification of said low-density heparan sulfate-proteoglycan by ion exchange chromatography followed by molecular sieve chromatography and fractionation in a density gradient.
A method for the immunological determination of low-density heparan sulfate-proteoglycan antigen of approximately 620-680 kd from basal membrane tissue of human placenta, or its antigenic determinants in a sample of body fluids, comprising the steps of:
- a) reacting the sample of antigen or its antigenic determinants with an amount of antibodies specific to said antigen that is in excess quantity relative to the sample being investigated to form an antigen-antibody complex and unreacted antibodies;
- and
- b) determining the presence or amount of antigen in the sample by immunological methods.
A substantially pure antibody preparation specific for heparan sulfate-proteoglycan from immunized animals containing antibodies against low-density heparan sulfate-proteoglycan of about 620-680 kd (.delta.=1.33--1.39) extracted from basal membrane tissue.
Substantially pure low-density heparan sulfate-proteoglycan of about 620-680 kd from human placenta basal membrane tissue.
Substantially pure antibody specific to low-density heparan sulfate-proteoglycan of about 620-680 kd from human placenta basal membrane tissue.
Dependencies are inferred from the claim text.