Detection of receptor-ligand interactions using surface plasmon resonance: model studies employing the HIV-1 gp120/CD4 interaction.
Surface plasmon resonance (SPR), a label-free, real time optical detection principle, has been investigated for its potential to detect and quantitate macromolecular ligand-ligate interactions. As model systems, the interactions of the HIV-1 envelope glycoprotein, gp120, and the monoclonal antibody L-71, with a soluble form of the T-cell receptor CD4 (sCD4), were investigated. In an effort to demonstrate potential analytical applications of this technology, operational characteristics of the SPR instrumentation (BIAcore, Pharmacia) including stability of the sensing surface and reproducibility in the measurement of such macromolecular interactions were investigated. In addition, the ability to detect and quantitate sCD4 directly from unfractionated cell culture supernatants, such as Streptomyces lividans, was investigated. The results demonstrate that SPR has potential in quantitating macromolecular interactions in both purified and crude samples and that the reproducibility in, and sensitivity of, such determinations is comparable to other techniques.
Brigham-Burke
M
M
SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
Edwards
J R
JR
O'Shannessy
D J
DJ
D000911:Antibodies, Monoclonal; D015374:Biosensing Techniques; D015704:CD4 Antigens / Q000378:metabolism*; D003470:Culture Media; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D008024:Ligands; D012031:Refractometry; D015203:Reproducibility of Results; D013057:Spectrum Analysis / Q000379:methods*; D013302:Streptomyces
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D003470:Culture Media; D015699:HIV Envelope Protein gp120; D008024:Ligands