Recombinant HIV-II Antigen
Certificate of Analysis and Data Sheet
Description: This antigen is an E.coli-derived recombinant protein that composes all of the reported immunogenic determinants found in gp39. The gene encoding this fusion protein was synthesized using codons optimized for E.coli expression and doesn’t represent a linear HIV-II envelope sequence. rHIV-II is a non-glycosylated, polypeptide chain, having a molecular mass of 16,127 Dalton. The rHIV-II is purified by standard chromatographic techniques.
Source: Escherichia Coli.
Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation & packaging:
The protein was lyophilized after extensive dialysis against
5mM-phosphate
buffer pH=7.3 containing 0.01% SDS.
Solubility: The lyophilized rHIV-II is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=5.17).
Purity: Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE
(Limit of acceptance:³98.0%. No more than 2% total impurities; no single impurity greater than 1%)
Stability: Lyophilized rHIV-II, although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHIV-II is best stored refrigerated at 4°C.
Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity: This antigen was thoroughly tested in various tests as well as ELISA using the standard BBI serum panels & 3 gp39-specific monoclonal antibodies. All antibody detection tests on this antigen were as sensitive or more than the standard Abott third generation tests.
Protein content: Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 3.14 as the extinction coefficient for
a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of
protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a calibrated solution of rHIV-II as a Reference Standard.
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