Advanced Developement of Leishmania Tropical Skin Test Antigen
Abstract
Leishmania tropica Skin Test Antigen ( LtSTA) is lysate of microfluidized L. tropica promastigotes. The product is manufactured at a protein concentration of 0.5 mg/mL. Lot consistency is established by densitometry readings of SDS-PAGE gels and delayed-type skin tests in sensitized guinea pigs. The dominant proteins in the promastigote lysate are found at 8 kDa, 20kDa, 30kDa, and 56-58 kDa; of these, the 8 kDa and 56-58 kDa entities are most prominent. The 8 kDa component is believed to be ubiquitin consisting of several proteases. The 56-58 kDa component was shown to contain metalloproteases, trypanothione reductase and dihydrolipamide dehydrogenase by NanoLC-ESI-MS/MS peptide sequencing technology. By densitometry, the relative amounts of the five major proteins in the lysate were as follows: 8 kDa (10.1-31%), 20 kDa (3.9-7.8 %), 30 kDa (3.4-9.2%), 56 kDa (28.9-40.1%), 58 kDa (31.3-40.1). Based on a validated assay using sensitized guinea pigs, lotscontaining 0.34 mg/mL to 0.61 mg/mL of protein can be considered equipotent to 0.5 mg/mL.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2011
- Accession Number
- ADA553653
Entities
People
- H. S. Nielsen Jr.