Figure 1

1D Demonstration of the depletion capabilities of the IgY-12 High Capacity SC Spin Column kit on patient serum. Depletion of patient serum was performed as indicated by manufacturer's instructions. Low and High abundant fractions were collected for each sample and run on a 1D 4-20% Tris-Gycine SDS-PAGE gel. A) Whole serum (lanes 2, 3) was incubated with the column containing antibodies against 12 of the high abundant serum proteins. Low abundant proteins (lanes 4, 5) were collected as the flowthrough, the column was washed (lanes 6, 7) and the high abundant proteins eluted (lanes 8, 9). Briefly the observed high abundant proteins were compared to the known sizes of the expected proteins as indicated. B) Equal volumes of serum from each of the six patients within each category (LTNP, HAART, and Negative) were pooled together to create a stock of each condition, independent of patient-to-patient variability. Twenty microliters of each stock was subjected to depletion and equal concentration of Low and High fraction were run on a 1D gel. Lanes 2, 3 and 4 are the low abundance fractions of the pooled LTNP, HAART, and Negative patients, respectively. Lanes 6, 8, and 10 are the high abundance fractions of the pooled LTNP, HAART, and Negative patients, respectively. The indicated arrows represent differentially expressed proteins that were excised, trypsinized, and identified using MALDI-TOF for preliminary protein screening.