Solid phase enzyme immunoassay of human interleukin 2 utilizing antibodies against synthetic IL-2 peptides.
In the present study, solid phase enzyme immunoassay utilizing antibodies against synthetic IL-2 peptides was used for quantitative measurements of human recombinant and lymphoid IL-2 preparations. The 27-peptide MCF-III-6 (Leu-Glu-His-Leu-Leu-Leu-Asp-Leu-Gln-Met-Ile-Leu-Asn-Gly-Ile-Asn-Asn-- Tyr-Lys-Asn-Pro-Lys-Leu-Thr-Arg-Met-Leu) that comprises the region 14-40 from the IL-2 amino acid sequence was synthesized and used for immunization of rabbits. Resulting anti-MCF-III-6 polyvalent rabbit antibodies reacted specifically in EIA up to dilution of 10(-7) with the MCF-III-6 peptide used for immunization as well as with 16-peptide I-16 (Cys-Nle-Gly-Ile-Asn-Asn-Tyr-Lys-Asn-Pro-Lys-Leu-Thr-Arg-Met-Leu) that comprises the region 27-40 from the IL-2 amino acid sequence. The anti-MCF-III-6 antibody reacted also with human recombinant IL-2 preparations obtained from three producers (Cetus, Riga and Amersham), to the concentration of 0.1 ng/ml, and with various human lymphoid IL-2 preparations. Direct correlation was observed between quantitative measurements of human lymphoid IL-2 by EIA and by CTLL bioassay. It can be concluded that utilization of synthetic IL-2 peptides provides a suitable and comparatively unexpensive immunogen for the production of IL-2 antibodies and that the solid phase EIA using such antibodies can be employed as a rapid, reproducible, and sensitive method for quantitative examination of both recombinant and lymphoid IL-2 preparations.
Jandlová
T
T
Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Praha.
Bubeník
J
J
Krchnák
V
V
Vágner
J
J
Voitenok
N N
NN
Gren
E
E
Símová
J
J
Mach
O
O
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis; D002460:Cell Line; D006801:Humans; D007124:Immunoenzyme Techniques; D007376:Interleukin-2 / Q000032:analysis* / Q000276:immunology; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D000906:Antibodies; D007376:Interleukin-2; D010455:Peptides; D011994:Recombinant Proteins