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P085
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P085
Abstracts / Human Immunology 75 (2014) 50–141
RAPID METHOD TO ELIMINATE HIGH BACKGROUND ON LUMINEX-SINGLE ANTIGEN BEADS (L-SAB) IN SERA FROM PATIENTS WITH VENTRICULAR ASSIST DEVICES (VAD). Ge Chen, Flavia Sequeira, Dolly Tyan. Stanford, Palo Alto, CA, United States. Aim: Antibody to albumin (Alb-Ab) can develop in VAD patients and causes high background on L-SAB. The aim of the study was to eliminate the non-specific signals by BSA coated bead (BK-Bead) absorption. Methods: BK-Bead absorption: Sera with high background on L-SAB (One Lambda, Inc.) were absorbed twice by mixing BK-Beads with serum at 1:10 ratio and vortexing for 10 min at RT each time. Absorbed sera were then retested by L-SAB per manufacturer’s protocol. Results: Alb-Ab developed 15–30 days after VAD implantation and significantly decreased within 1– 3 months after transplantation in all VAD patients (Fig. 1). A total of 77 samples from 29 patients were tested by L-SAB before and after BK absorption. BK absorption successfully eliminated the Alb-Ab high background on #1 NC bead as well as HLA-Ag specific beads in all VAD samples with high background (Fig. 2). BK-Bead absorption has no effect on the detection of true HLA-Ab in non-VAD samples (Fig. 3). Conclusion: This simple and rapid BK-Bead absorption procedure can efficiently remove Alb-Ab from VAD patient samples and eliminate its non-specific signals without affecting the detection of true HLA-Abs on LSAB.
Fig. 1. LMX-SAB negative control beads follow-up.
Abstracts / Human Immunology 75 (2014) 50–141
Fig. 2. Effect of BK ansorption on MFI values of LMX-SAB negative control beads in VAD samples.
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P085
Abstracts / Human Immunology 75 (2014) 50–141
RAPID METHOD TO ELIMINATE HIGH BACKGROUND ON LUMINEX-SINGLE ANTIGEN BEADS (L-SAB) IN SERA FROM PATIENTS WITH VENTRICULAR ASSIST DEVICES (VAD). Ge Chen, Flavia Sequeira, Dolly Tyan. Stanford, Palo Alto, CA, United States. Aim: Antibody to albumin (Alb-Ab) can develop in VAD patients and causes high background on L-SAB. The aim of the study was to eliminate the non-specific signals by BSA coated bead (BK-Bead) absorption. Methods: BK-Bead absorption: Sera with high background on L-SAB (One Lambda, Inc.) were absorbed twice by mixing BK-Beads with serum at 1:10 ratio and vortexing for 10 min at RT each time. Absorbed sera were then retested by L-SAB per manufacturer’s protocol. Results: Alb-Ab developed 15–30 days after VAD implantation and significantly decreased within 1– 3 months after transplantation in all VAD patients (Fig. 1). A total of 77 samples from 29 patients were tested by L-SAB before and after BK absorption. BK absorption successfully eliminated the Alb-Ab high background on #1 NC bead as well as HLA-Ag specific beads in all VAD samples with high background (Fig. 2). BK-Bead absorption has no effect on the detection of true HLA-Ab in non-VAD samples (Fig. 3). Conclusion: This simple and rapid BK-Bead absorption procedure can efficiently remove Alb-Ab from VAD patient samples and eliminate its non-specific signals without affecting the detection of true HLA-Abs on LSAB.
Fig. 1. LMX-SAB negative control beads follow-up.
Abstracts / Human Immunology 75 (2014) 50–141
Fig. 2. Effect of BK ansorption on MFI values of LMX-SAB negative control beads in VAD samples.
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