HUMAN SERUM FOLATE CAN BE MEASURED USING FOLATE BINDING PROTEIN LINKED TO ENZYME-LABELED PROTEIN LIGAND BINDING ASSAY (ELPLBA) AS WELL AS ELISA

Muhamad Arif Budiman, MAB (2018) HUMAN SERUM FOLATE CAN BE MEASURED USING FOLATE BINDING PROTEIN LINKED TO ENZYME-LABELED PROTEIN LIGAND BINDING ASSAY (ELPLBA) AS WELL AS ELISA. Acta Biochimica Indonesiana, 1 (2). pp. 59-67. ISSN 2654-6108

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Abstract

Background: Folate is an important substance used for purine and pyrimidine nucleotide synthesis. One measurement of folate that already establishes is using ELISA (Enzymelinked immunosorbent assay) method. Folate binding protein is a protein that can bind folate,therefore it considered can be used as a tool that can replace antibody dependent ELISA method. Objectives: The aim of this research was to create a method for folate measurement in serum called Enzyme-labeled protein ligand binding assay (ELPLBA) by replacing antibody as used in ELISA method with folate binding protein (FBP) that purified from the whey of milk. Methods: The method is tested using 20 serum samples and compared to ELISA. Folate binding protein was purified from bovine's milk using ammonium sulfate up to 90% saturated, DEAE-cellulose anion exchange chromatography and affinity chromatography. SDS-PAGE and western blot were used to establish the protein band of FBP that has molecular weight of ~25-35 kDa. ELPLBA was arranged with stationary phase using aminohexyl-agarose, and folic acid linked on it using carbodiimide. Results: The result show there was no significant difference of folate concentration between ELPLBA (14.804 ± 2.795) and ELISA method (13.859 ± 3.638), p = 0.363. Conclusion: ELPLBA method show similarity for determination of folate in serum which was the same as standard folate measurement (ELISA).

Item Type:	Article
Subjects:	Q Science > QD Chemistry R Medicine > R Medicine (General)
Divisions:	Fakultas Kedokteran > Kedokteran
Depositing User:	Muhamad Arif Budiman
Date Deposited:	09 Sep 2021 09:12
Last Modified:	09 Sep 2021 09:12
URI:	http://repository.uhamka.ac.id/id/eprint/11090

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