A novel strategy for efficient production of anti-V3 human scFvs against HIV-1 clade C

BMC Biotechnology

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).

ISSN: 1472-6750