Recombinant HIV-I Antigen

( rHIV-I )

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 gp41 and a small portion of gp120. The gene encoding this fusion protein was synthesized using codons optimized for E.coli expression and doesn’t represent a linear HIV-I envelope sequence.

rHIV-I is a  non-glycosylated, polypeptide chain, having a molecular mass of 27,333 Dalton .

The rHIV-I 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-I is soluble in water and most aqueous buffers below and above the isoelectric point (pI=9.7).

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-I, although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHIV-I is best stored refrigerated at 4°C.

 

Biological Activity:

This antigen was thoroughly tested in various tests as well as ELISA using the standard BBI serum panels.

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 2.94 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 HIV-I as a Reference Standard.

Usage: This material is offered  for research, laboratory or further manufacturing purposes.

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