Complete freund s adjuvant cfa

A CH401 peptide, which includes the epitope sequence of the anti-human epidermal growth factor receptor 2 (HER2) mAb, was determined using multiple antigen peptides (MAP) with a partial amino acid sequence of HER2/neu previously (34) . The peptide was synthesized using a Rink amide resin (0.4-0.7 mmol/g), an ACT357 peptide synthesizer (Advanced Chemtech, Louisville, KY, USA) and a multiple antigen peptide named CH401MAP, a 20-mer peptide of the HER2 molecule that was synthesized as an antigen peptide. The HER2 peptide was emulsified with complete Freund's adjuvant (CFA) (Wako Pure Chemical Industries, Ltd, Osaka, Japan) (50 μg/head, 100 μL, 1:1, v/v) and administered intraperitoneally to the PBMC-transplanted NOG-hIL-4-Tg mice in which the hIL-4 level was greater than 100 pg/mL in the plasma. For the negative control, an equal volume of PBS was emulsified and injected into PBMC-transplanted NOG-hIL-4-Tg mice. Boosters were performed using incomplete Freund's adjuvant (IFA) (Wako Pure Chemical Industries, Ltd) at 2 weeks after the first immunization. Two weeks after the booster, the mice were sacrificed for analyses.