Johnson H M, Langford M P, Lakhchaura B, Chan T S, Stanton G J
J Immunol. 1982 Dec;129(6):2357-9.
A synthetic peptide corresponding to the N-terminal amino acid sequence of human gamma-interferon (HuIFN gamma), based on the cDNA sequence, was used to produce antibodies in rabbits that were reactive with native HuIFN gamma. Antibodies from all immunized rabbits neutralized the antiviral activity of HuIFN gamma. Significant neutralization of other HuIFN and mouse IFN was not observed. The peptide had the sequence Cys-Tyr-Cys-Gln-Asp-Pro-Tyr-Val-Lys-Glu-Ala-Glu-Asn-Leu-Lys-Lys-Tyr-Phe-Asn-Ala ,and was coupled to keyhole limpet hemocyanin by disulfide linkage with the use of cystamine. The specificity of the antibodies produced to the peptide was compared to that of antibodies produced to native HuIFN gamma by neutralization of HuIFN gamma and by reactivity with peptide in the enzyme-linked immunosorbent assay (ELISA). The ratio of anti-peptide antibody neutralization of HuIFN gamma vs reactivity with peptide in the ELISA was at least 28-fold lower than for anti-HuIFN gamma antibody. Thus the antibodies to peptide and to HuIFN gamma were directed primarily against different determinants on native HuIFN gamma or the anti-HuIFN gamma antiserum probably contained antibodies to additional determinants. The anti-peptide antibodies should be useful for further characterization and purification of HuIFN gamma.
基于cDNA序列，合成了一段与人类γ干扰素（HuIFNγ）N端氨基酸序列相对应的肽段，并用其在兔体内产生与天然HuIFNγ反应的抗体。所有免疫兔的抗体均中和了HuIFNγ的抗病毒活性。未观察到对其他HuIFN和小鼠IFN的显著中和作用。该肽段序列为Cys-Tyr-Cys-Gln-Asp-Pro-Tyr-Val-Lys-Glu-Ala-Glu-Asn-Leu-Lys-Lys-Tyr-Phe-Asn-Ala，通过胱胺利用二硫键与匙孔血蓝蛋白偶联。通过中和HuIFNγ以及在酶联免疫吸附测定（ELISA）中与肽段反应，比较了针对该肽段产生的抗体与针对天然HuIFNγ产生的抗体的特异性。抗肽抗体对HuIFNγ的中和作用与在ELISA中与肽段反应性的比值比抗HuIFNγ抗体至少低28倍。因此，针对肽段和HuIFNγ的抗体主要针对天然HuIFNγ上不同的决定簇，或者抗HuIFNγ抗血清可能包含针对其他决定簇的抗体。抗肽抗体应有助于对HuIFNγ进行进一步的表征和纯化。
