Human immunodeficiency virus: novel enzyme-linked immunoassays for quantitation of envelope glycoprotein 120.
Two novel enzyme-linked immunoassays (ELISA) for the quantitation of human immunodeficiency virus type 1 (HIV-1) coded glycoprotein with an Mr 120 (gp120) are described. These are based on the highly specific interaction between gp120 and the mannose-specific lectins from Narcissus pseudonarcissus (NPL) and Galanthus nivalis (GNL). Two systems were developed: (1) an HIV-protein ELISA using HIV-protein (also containing HIV-gp120) for the solid phase and NPL as a detector and (2) a lectin-ELISA using the NPL bound to the solid phase and GNL as detector. The HIV-protein ELISA was validated for quantitation of gp120 within the range 3 to 600 ng/ml; the lectin-ELISA for concentrations between 0.6 and 20000 ng gp120/ml. Serum components did not interfere with the binding of gp120 to the lectins. The ELISAs were used for the quantitation of gp120 in HIV-infected CEM cells in vitro. It was found that gp120 appeared in the medium earlier after infection than HIV-p24 and reverse transcriptase, suggesting that gp120 is released as free glycoprotein. Moreover, the ELISAs were also applied successfully for the detection of compounds that bind to gp120 and for the identification of antibodies directed against the highly pathogenic mannan portion of gp120. These ELISAs are considered to be suitable also for the detection of gp120 in the serum of HIV-infected individuals.
Weiler
B E
BE
Institut für Physiologische Chemie, Abteilung, Angewandte Molekularbiologie Universität Mainz. F.R.G.
Schäcke
H
H
Bachmann
M
M
Brigido
L
L
Gilbert
M
M
Mills
J
J
Matthes
E
E
Forrest
J M
JM
Müller
W E
WE
D000818:Animals; D002138:Calibration; D002241:Carbohydrates / Q000276:immunology; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D027801:Galanthus; D005872:Giardia / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000097:blood* / Q000276:immunology; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000378:metabolism; D037121:Plant Lectins; D012680:Sensitivity and Specificity
D002241:Carbohydrates; D015699:HIV Envelope Protein gp120; D037102:Lectins; D037121:Plant Lectins