A micromethod for the isolation of brush border membrane fragments from single peroral duodenal biopsies, and their subsequent analysis by polyacrylamide gel electrophoresis is described. The quantity of biopsy material used varied between 5 and 15 mg wet weight, leaving enough mucosa for histological examination. By cutting the gels longitudinally into two halves it was possible to identify several maltases, sucrase, isomaltase and lactase and to correlate these enzymatic activities with distinct co-migrating protein peaks. For alkaline phosphatase and enterokinase this correlation was not possible. This method is suitable for the study on single biopsies of the molecular alterations occurring in the various congenital enzyme deficiencies of the human small intestine.