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. 2004 Apr 27;90(10):2032–2041. doi: 10.1038/sj.bjc.6601825

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Copyright © 2004 Cancer Research UK

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Analysis of the Ab3 anti-anti-Id scFv 69 response in sera of BALB/c mice (immunisation protocol 1) by inhibition of the binding of anti-Id scFv 69 (Ab2) on immobilised trastuzumab F(ab′)₂ fragments (Ab1) by inhibition ELISA. Serial dilutions of preimmune sera or sera from mice each of the three groups: primed with HER-2/neu ECD-Fc fusion protein or with PBS or with anti-Id scFv 69 were preincubated with soluble anti-Id scFv 69. This solution was subsequently incubated for 2 h on trastuzumab F(ab′)₂ fragments followed by the detection of bound scFv by HRP-conjugated M2 anti-FLAG mAb. The results obtained are expressed as percent inhibition at each serum dilution.