Table 2

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

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