Methods for protecting silica sorbents used in high-performance liquid chromatography from strongly adsorbed impurities during purification of human recombinant insulin.
One of the main stages of human recombinant insulin (HRI) production is the hormone purification by means of reversed phase high-performance liquid chromatography (RP HPLC). The optimization of this stage determines the increase of the total manufacturing yield. Therefore, the cost of the sorbent used in HPLC influences the cost of the manufacturing product, i.e. HRI substance. However, resolution between HRI and its admixtures decreases with time. The reason for this is the sorbent contamination with strongly adsorbed impurities (SAI) which are accumulated during elution. In the following research several methods for sorbent protection are studied. The opinion that SAI are mainly high-molecular weight compounds was examined using gel filtration. Different sorbent types were tested for the use in guard column. The results obtained were applied and improved at preparative level.
Gusarov
D
D
Experimental Biotechnological Plant, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Mikloukho-Maklaya st., 16/10, Moscow 117997, Russian Federation. gusarovda@list.ru
Lasman
V
V
Bayramashvili
D
D
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry*
D007328:Insulin; D011994:Recombinant Proteins; D012822:Silicon Dioxide