Biosynthetic human

Biosynthetic Human Insulin from E. coli. Insulin [9004-10-8] a polypeptide hormone, stimulates anaboHc reactions for carbohydrates, proteins, and fats thereby producing a lowered blood glucose level. Porcine insulin [12584-58-6] and bovine insulin [11070-73-8] were used to treat diabetes prior to the availabiHty of human insulin [11061 -68-0]. AH three insulins are similar in amino acid sequence. EH LiHy s human insulin was approved for testing in humans in 1980 by the U.S. EDA and was placed on the market by 1982 (11,12). 

Fig. 1. (a) Process flow sheet for human insulin production, recovery, and purification (12) (b) corresponding steps in recovery of biosynthetic human... 

Nielsen, R. G., Riggin, R. M., and Rickard, E. C., Capillary zone electrophoresis of peptide fragments from trypsin digestion of biosynthetic human growth hormone, /. Chromatogr., 480, 393, 1990. 

Although gradient elution is generally required for RPLC separations of proteins, isocratic elution can be successful in some instances. For example, isocratic elution has been used for the determination of purity of production batches of biosynthetic human growth hormone (HGH).42 The method was used to... 

Figure 4.12 Analysis of biosynthetic human growth hormone by isocratic reversed-phase chromatography. To generate all possible degradation products, a production lot of HGH was exposed to 40°C. The profile of the unstressed HGH is shown in the upper trace. (Reproduced from R.M. Riggin, G.K. Dorulla, and DJ. Miner, Anal. Biochem., 167 199 [1987]. With permission from Elsevier Science.)...

Kroeff, E. et al. (1989). Production scale purification of biosynthetic human insulin by reverse phase high performance liquid chromatography. J. Chromatogr. 461, 45-61. 

Biphasic insulins are fixed dose combinations of a short-acting and intermediate-acting insulin in various proportions. Examples are Humulin, a biosynthetic human insulin (rDNA origin) suspension with respectively 20%, 30% and 40% regular and 80%, 70% and 60% isophane insulin and Mixtard, a biphasic biosynthetic human insulin suspension with respectively 10%, 20% and 40% soluble and 90%, 80% and 60% isophane insulin. 

Human forms are derived from recombinant or biosynthetic human insulin. Animal sources are derived from purified pork insulin. 

Other problems that may be encountered are related to the immunologic effects of insulin use. Certain forms of insulin may evoke an immune reaction and stimulate antibody production. These anti-insulin antibodies may cause an allergic reaction in some individuals, as well as a resistance to the exogenous insulin molecule. As discussed previously, the incidence of these immunologic reactions seems to be greater when animal (i.e., pork) forms of insulin are used. Consequently, these problems are often resolved by switching the patient to another type of preparation, preferably biosynthetic human insulin. 

R. M. Riggin, C. J. Shaar, C. K. Dorulla, D. S. Lefeger, and D. J. Miner, High-performance-exclusion chromatographic determination of the potency of biosynthetic human growth products, J. Chromatogr., 435 301 (1988). 

Performance parameters were established for the use of capillary zone electrophoresis (CZE) in the characterization of biosynthetic human growth hormone (hGH). 

The observed precision is comparable to the values we previously reported for biosynthetic human insulin (16). It also is similar to independent results obtained using a totally automated system (2.9% RSD) and much better than that reported for manual injection (11.8% RSD), both using a hydrodynamic injection technique (21). Finally, the observed precision for the percent desamido, which is really an area ratio similar to what would be obtained by comparison to an internal standard, is excellent for the 10-nL or larger injections. Although the data are insufficient to make a definitive conclusion, it suggests that the observed error is comparable to that obtained from many chromatographic techniques. It also suggests that one of the predominant sources of error is imprecision in the injection volume. The error in injection volume was recently characterized (19). They also reported approximately 1-3% RSD in peak areas for vacuum injection of various compounds. 

The establishment of a physicochemical potency assay for biosynthetic human growth hormone provides an interesting example of an approach for validation of such assays. The hypophysectomized rat growth promoting assay used tradionally has a precision of approximately 15-20% RSD, which is clearly not adequate for controlling well-characterized products. In order to select a suitable assay to replace the in-vivo bioassay, the biopotencies... 

Chance RE, Frank BH. Research, development, production, and safety of biosynthetic human insulin. Diabet Care 1993 16(Suppl 3) 133 i2. 

Baker, R. S., Schmidtke, J. R., Ross, J. W., and Smith, W. C. (1981). Preliminary studies on the immunogenicity and amount of Escherichia coli polypeptides in biosynthetic human insulin produced by recombinant DNA technology. Lancet 2(8256), pp. 1139-1142. 

Petrides, D., Sapidov, E., and Calandranis, J. (1995). Computer-aided process analysis and economic evaluation for biosynthetic human insulin production—A case study. Biotechnol. Bioeng. 48, 529-541. 

B. Example 5 An Economic Analysis of Biosynthetic Human Insulin Production... 

Ilium, L., Faraj, N., Davis, S., Johansen, B., and O Hagan, D. (1990), Investigation of the nasal absorption of biosynthetic human growth hormone in sheep Use of a bioadhesive microsphere delivery system, Int. J. Pharm., 63,207-211. 

Accurate measurement of proinsulin has been difhcult for several reasons the blood concentrations are low antibody production is difficult most antisera cross-react with insulin and C-peptide, which are present in much higher concentrations the assays measure intermediate cleavage forms of proinsulin and reference preparations of pure proinsulin are not readily available. However, a more sensitive nonequiUb-rium RIA method for measuring proinsiilin was developed by adsorbing the initial antiserum with biosynthetic human C-peptide coupled to agarose to eliminate cross-reactivity with C-peptide.An enzyme-linked immunosorbent assay (ELISA) has been described that employs an antibody to C-peptide as the coating antibody and antiinsulin antibody for detection. The detection limit is 0.25 pmol/L. ... 

Bowsher R, Apathy J, Ferguson A, Riggin RM, Henry DP. Cross-reactivity of monomeric and dimeric biosynthetic human growth hormone in commercial immunoassays. CHn Chem 1990 36 362-6. 

Rougeot C, Marchand P, Dray E, et al. Comparative study of biosynthetic human growth hormone immunogenicity in growth hormone deficient children. Horm Res 1991 35 76-81. 

HUMULIN R REGULAR U-500 (concentrated insulin human injection, USP) [/s call expressed biosynthetic human insulin] Eli Lilly 20 mL vials of zinc-insulin crystals dissolved in a clear fluid 500 Units/mL 100 units/ mL (U-100) 0.24 mg sodium citrate 1.04 mg citric acid monohydrate 9.6 mg mannitol 0.8 mg polysorbate 80 16mg/mL glycerin, 2.5mg/mL m-cresol as a preservative 0.085 mg/mL (0.017 mg/ 100 units) zinc-oxide (sodium hydroxide and/or hydrochloric acid may be added during manufacture to adjust the pH)... 

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