Human growth hormone recombinant preparations

Table 8.8. Recombinant human growth hormone (rhGH) preparations approved for general medical...

GH assays now use mouse monoclonal antibodies, and some of these assays are able to discriminate GH variants. Most procedures use recombinant-derived GH for labeling with a tracer and calibration material. The latter is usually prepared gravimetricaUy and verified by comparison with an international reference preparation (IRP), such as the World Health Organization s (WHO s) international standard, IRP 80/505 human growth hormone recombinant (hGHr), which has a potency of 3.3IU/mg of r-hGH, or other standard preparations, such as WHO IRP 66/217 or 88/624. The assay diluent, however, can vary considerably from one assay to another and is a potential source of bias some commercial kits use phosphate-buffered saline, and others use horse, calf, or human serum. 

FIGURE 2.10 Analytical SEC for determination of abnegates in preparations of recombinant human growth hormone using Superdex 75 HR 10/30. [Reproduced from Hagel (1993). with permission.]... 

There are many examples of ELIS As used for detecting host cell impurities in the literature. Pauly et al.12 developed an ELISA to detect impurities in erythropoietin that had a detection limit of around 0.05 ng/ml. SDS polyacrylamide gel and Western blot analysis were used to confirm the spectrum of proteins detected and to demonstrate the specificity of the antibody preparation. Anicetti et al.14 describe an assay for the detection of E. coli proteins in recombinant DNA-derived human growth hormone. Whitmire and Eaton15 report on an immuno-ligand assay for quantitation of process-specific E. coli host cell contaminant proteins in a recombinant bovine somatotropin. 

Figure 8.14. Production of recombinant human growth hormone (rhGH) in E. coli (as an intracellular protein). Subsequent to fermentation, the cells are collected by centrifugation or filtration. After homogenization, nucleic acids and some membrane constituents are precipitated by the addition of polyethyleneimine. Ammonium sulphate precipitation of the supernatant concentrates the crude rhGH preparation. Chromatographic purification follows, as illustrated...

Human growth hormone is produced by recombinant DNA technique. The preparation available are ... 

Samples included rCD4 (3), rCD4-IgG (4), rgpl20 (5), rtPA (Activase ) and rhuMAb HER2 (6) all of which were purified from transfected Chinese hamster ovary (CHO) cells. Human rtPA from transfected 293 (human embryonic kidney) cells was prepared as described (7). Bowes melanoma tPA was obtained from American Diagnostica. Human plasma factor IX was kindly provided by Kenneth J. Smith (8). Recombinant human growth hormone (Protropin ), isolated from transfected E. coli (10), was included as a negative control. 

Recombinant human tumor necrosisb factor alpha from an extract of Escherichia coli Recombinant human growth hormone preparation (rhGH)... 

Recombinant human growth hormone preparation (rhGH)... 

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