|Screening of EBV transformed PBMCs isolated from a drug naïve HIV-1 infected patient (# 254) for V3 reactivity|
|1Total number of PBMCs isolated (millions) from HIV-1 infected patient # 254||4.8|
|2Number of cells plated per well||100000|
|3Total number of wells plated with the cells and EBV transformed in 96 well culture plate||48|
|4Number of wells successfully EBV transformed||48|
|5Number of wells secreting anti-V3 Abs (OD>2) in 96 well stage(first screening)||7|
|6Number of wells secreting anti-V3 Abs (OD>0.6) in 24 well stage(second screening)||5|
|7OD at flask stage(Final screening) (OD>1.0)|
1 PBMCs were isolated from a HIV positive drug naïve patient sample (# 254). A total of 4.8 million PBMCs were isolated.
2 Number of cells plated per well was 100000.
3 Forty eight wells were subjected to EBV transformation.
4 All the 48 wells were successfully transformed (transformation efficiency 100%).
5 7/48(approx 14.6%) wells showed positive binding (OD>2) with V3 peptide and expanded to 24 well stage.
6 5/7 (approx 71%) wells showed positive binding (OD>0.6) with V3 peptide and pooled down and grown in T-25 flask.
7 Final screening for anti-V3 antibody binding reactivity was done at flask stage (OD>1.0).
Kumar et al.
Kumar et al. BMC Biotechnology 2012 12:87 doi:10.1186/1472-6750-12-87