WORLD JOURNAL OF ADVANCE
HEALTHCARE RESEARCH

Dezheen Shabaan Kucher, Thikra Ali Allwsh*

ABSTRACT

Alpha-Klotho (α-Klotho) is considered of the key regulatory proteins involved in maintaining homeostasis in biological systems, particularly in neurons and neurotransmitters. The present study aimed to isolate and partially purify the α-Klotho protein from the plasma using various biotechnological processes, as well as finding the molecular weight of α-Klotho. The research involved using plasma from a healthy person for partial purification of α-Klotho by using three sequential steps: Saline displacement precipitation, dialysis, and size exclusion chromatography were used with Sephadex gel number G-75. The results showed an increase in the specific concentration and the number of isolations of α-Klotho (22.74) (1.3), respectively? following first step in precipitation and the recovery was 87% of α-Klotho. Also, the specific concentration increased to (33.13) in dialysis. The recovery amount of the total α-Klotho level reached 82%. In addition, two protein peaks (A and B) were detected, with α-Klotho corresponding to peak B, showing a purification fold of 38.5 compared with the crude plasma after the gel filtration process, with a 71-recovery rate. Electrophoretic (SDS-PAGE) was performed to confirm the purity of the α-Klotho, and its molecular weight was determined by both gel filtration and SDS-PAGE techniques, revealing an approximate molecular weight of 68 and 66 kDa, respectively.