Human plasma derived drugs separation by fractionation of plasma with polyethylene glycol

There are variety of purification techniques for separation of human plasma proteins such as salting out, ion exchange chromatography and ethanol fractionation. There are limitations for each method, for example in salting out method, the salt has to be removed in an additional step. Ion exchange chromatography is difficult for scaling up, and plasma fractionation is a time consuming method and it needs machinery and plants. In the present study the fractionation of human plasma by polyethylene glycol was investigated. The purpose of this study was to investigate the possibility of the fractionation of human plasma by polyethylene glycol. Human plasma fractionation was carried out by using polyethylene glycol with different concentrations from five to twenty percent, followed by centrifugation. After centrifugation the supernatant was used for further fractionation by addition of a higher concentration of polyethylene glycol. Suitable intermediate sources for protein purification were obtained by fractionation of human plasma by polyethylene glycol. Fibrinogen in fraction 5%, IgG and IgM in fraction 10%, IgA in fraction 20%, and finally albumin and alpha1- Antitrypsin in supernatant 20% of polyethylene glycol were achieved. By our study we could obtain four different fractions as intermediate sources for protein purification which cannot be easily obtained from plasma fractionation by cold ethanol fractionation

[ preparation of albumin as a biological drug from human plasma by fiber filtration]

In recent years, consumption of whole-blood for the treatment of patients has decreased but use of biological plasma-derived medicines such as albumin, immunoglobulin and coagulation factors have increased instead. Paying attention to albumin molecular structure is important for its isolation from human plasma. Albumin is a single-chain protein consisting of about 585 amino acids and a molecular weight of 66500 Daltons. Albumin is a stable molecule and it is spherical in shape. There are different methods for human albumin preparation. Considering the large consumption of this biological drug in clinical settings, methods with fewer steps in production line are of big advantage in saving time and manufacturing more products. In this project, we prepared human albumin using hollow fiber cartridges in order to omit the rework on fraction V+VI. Human albumin is usually produced by the application of cold ethanol method, where albumin is obtained from fraction V by doing a rework on fraction V+VI to separate fraction V. In the current work, human albumin was prepared from fraction V+VI by the help of hollow fiber cartridges. With a concentration of 20%, the obtained albumin had 96.5% of monomer and 3.5% of polymer and polymer aggregate. Comparing the obtained human albumin with a number of commercial human albumin samples by the use of SDS-page, the results were satisfactory regarding the 3.5 percent polymer and aggregate rate for the prepared albumin

New synthesis of aminorhodanin and condensed derivatives

Aminorhodanins are heterocyclic compounds of dithiocarbazoyl which include sulphur and nitrogen in their structures. Different research studies have been performed on rhodanin and it-s condensed derivatives and several reports have been issued indicating their antimicrobial properties. In the present work, first a new method has been developed for production of aminorhodanin, which include one more nitrogen atom in third position compare to rhodanins molecule, then condensed reactions with aldehydes have been investigated. The probable biological properties of produced compounds will be investigated in another research study

Pasteurization of IgM-enriched immunoglobulin

Human plasma proteins are important for therapy or prophylaxis of human diseases. Due to the preparation of human plasma proteins from human plasma pools and risk of contamination with human viruses, different viral reduction treatments such as: pasteurization, solvent/detergent, dry heat treatment, steam treatment, beta-propiolactone/UV and nanofiltration have been implemented. As pasteurization can be performed for liquid protein, this method [a 10-hour heat treatment of the aqueous solutions at 60°C] was introduced into the manufacturing procedure of IgM-enriched immunoglobulin, to improve its safety further. The efficiency of this method for inactivation of viruses was evaluated by the use of Foot-and-Mouth Disease Virus [a non-enveloped virus] and Infectious Bovine Rhinotracheitis [IBR] Virus [a lipid-enveloped virus]. Pasteurization inactivated Foot-and-Mouth Disease Virus by 7 log10 and for IBR Virus by 5log10. These findings show a significant added measure of virus safety associated with pasteurization of IgM-enriched immunoglobulin preparation

Solvent-detergent treatment of IgM-enriched immunoglobulin

Viral safety of human plasma products plays a key role in their safe uses. Solvent- detergent [SD] virus-inactivation method has gained widespread popularity in the manufacture of biological products. This treatment which inactivates lipid-enveloped viruses effectively consists of incubation of a plasma protein solution in the presence of a non-volatile organic solvent and a detergent. In this study, IgM-enriched immunoglobulin was incubated at 24 °C for 6 h under slow stirring in the presence of tri[n-butyl] phosphate [0.3% w/w] as solvent and tween 80 [1% w/w] as detergent. After completion of the inactivation process and removal of the solvent-detergent, the ability of SD-treatment to remove Infectious Bovine Rhinotracheitis [IBR] virus [a lipid-enveloped virus] and Foot-and-Mouth Disease virus [a non-enveloped virus] were evaluated by "virus spiking studies" using a scaled down process. Reduction factor of 4 log was obtained for the SD-treatment of IgM-enriched immunoglobulin spiked with IBR virus. No virus inactivation was observed in the SD-treated IgM-enriched immunoglobulin, spiked with Foot-and-Mouth Disease virus. It was concluded that treatment of IgM-enriched immunoglobulin with TNBP-TWEEN 80 may be considered as an efficient lipid-enveloped virus inactivation step in the manufacture of this product