HUMAN SERUM ALBUMIN ISOLATION FROM UNUSED BLOOD PLASMA USING MODIFICATION OF KISTLER & NITSCHMANN METHOD

• Main Author: SETIYAWAN (NIM : 21115015), AGUS
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/20826
• Institution: Institut Teknologi Bandung

Human Serum Albumin (HSA) demand for therapy in Indonesia is very high and increasing every year, and it is met from imports. PT Bio Farma (Persero) as a life science company in Indonesia is currently developing blood product, one of which is HSA. Mastery of technology, methods and process optimization to produce high quality HSA independently is the main thing that will be done by Bio Farma. The utilization of the unused donor blood also provides a double benefit, as a source of raw material for HSA production and reduce the cost of blood waste treatment by PMI. The aims from this research is isolating HSA from unused blood plasma using modification of Kistler & Nitschmann method, to produce HSA that have same quality with commercial HSA. Initially, a design of experiment (DoE) with Response Surfaces Methode (RSM) was performed to determine the parameters most influential in albumin fractionation. The isolation of albumin was performed by the Kistler-Nistchmann method, and the results were evaluated for process optimization. The total protein content was tested by bicinchonic acid (BCA) protein test. Albumin purity was observed using sodium dodesil sulfat polyacrylamide gel electroforesis (SDS PAGE), followed by densitometry analysis using TotalLab program. The HSA native forms of samples and commercial HSA were compared with Native PAGE. The most influential parameters for albumin fractionation were pH (p-value 0.037), followed by ethanol concentration (p-value 0.064) and process temperature (p-value 0.228). The HSA in supernatant 2 was obtained by Kistler-Nistchmann method, have the same SDS PAGE and Native PAGE spectra profiles as the commercial HSA, with a mean yield of 48,4±13,3% w/w and purity of 78,2±1,4% (purity of commercial HSA 80,0±5,2%). This HSA was obtained from 2 fractionation stages, shorter than original Kistler-Nistchmann method that used 3 fractionation phases. It is concluded that the HSA in supernatant 2 can be used as intermediete product for HSA production that has the same quality as commercial HSA.