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Recombinant human luteinizing hormone: a partial physicochemical, biological and immunological characterization
J A Talbot1, R Mitchell, A M Hoy
1University of Manchester, Department of Medicine (Clinical Biochemistry), UK.
Molecular Human Reproduction
|October 1, 1996
Summary
The Serono recombinant human luteinizing hormone (rhLH) preparation exhibits higher in-vitro bioactivity and a more basic charge profile compared to other tested rhLH and pituitary preparations. Carbohydrate complexity was similar across all samples.
Area of Science:
- Biochemistry
- Endocrinology
- Glycobiology
Background:
- Recombinant human luteinizing hormone (rhLH) is crucial for reproductive health and requires thorough characterization.
- Understanding glycoform composition, including charge and carbohydrate structures, is essential for assessing rhLH efficacy and safety.
Purpose of the Study:
- To partially characterize the glycoform composition of a Serono rhLH preparation (LHadi).
- To compare the charge (pI) and internal carbohydrate complexity of Serono rhLH with a Crystal Chem rhLH and a pituitary International Reference Preparation (IRP).
Main Methods:
- Preparations were separated by charge using chromatofocusing and by carbohydrate complexity using concanavalin A (Con-A) lectin affinity chromatography.
- Quantification of luteinizing hormone (LH) was performed using immunoradiometric assay (IRMA) and Leydig cell bioassay (B-LH), calibrated against IRP 80/552.
Main Results:
- Serono rhLH demonstrated the highest in-vitro biopotency (18187 IU/mg) and biological:immunological ratio (1.99).
- Median pI values for bioactive LH were 5.5 (Serono), 5.52 (Crystal Chem), and 4.97 (pituitary IRP).
- All preparations contained <1% complex carbohydrate structures, with 36-44% intermediate and 56-63% simple forms of bioactive LH.
Conclusions:
- The Serono rhLH preparation exhibits superior in-vitro bioactivity and a more basic charge profile compared to Crystal Chem rhLH and pituitary IRP.
- Despite differences in bioactivity and charge, the carbohydrate moiety complexity appears similar across the evaluated rhLH and pituitary preparations.