Natural Pig Plasma Immunoglobulins Have Anti-Bacterial Effects: Potential for Use as Feed Supplement for Treatment of Intestinal Infections in Pigs

Hedegaard, Chris J.; Strube, Mikael L.; Hansen, Marie B.; Lindved, Bodil K.; Lihme, Allan; Boye, Mette; Heegaard, Peter M. H.

Generally, for all ELISAs washing steps consisted of four times washes in PBS with 0.05% Tween 20 (PBS-T). PBS-T with 1% bovine serum albumin (Sigma-Aldrich) was used as both blocking and dilution buffer. Coating was done with 100 μl/well at 4°C overnight. Blocking was done with 200 μl blocking buffer shaking for 30 min. at room temperature. Plates were 96 wells flat bottom Maxisorp plates (NUNC, Thermo Scientific, Denmark) and all operations were performed at room temperature unless specified otherwise. Development of plates was done by TMB Plus substrate (Kem-En-Tec, Taastrup, Denmark), 100 μl/well, and stopping colour development by 100 μl 0.5 M H 2 SO 4 (VWR-Bie & Berntsen A/S). Optical densities of microtiter plates were measured by Thermo Scientific Multiscan EX microplate reader at 450 nm absorbance and the background absorbance at 650 nm was subtracted.