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Growth inhibition assay (GIA)

PP Palak N. Patel
TD Thayne H. Dickey
CH Christine S. Hopp
AD Ababacar Diouf
WT Wai Kwan Tang
CL Carole A. Long
KM Kazutoyo Miura
PC Peter D. Crompton
NT Niraj H. Tolia
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GIA was performed as described in the protocol of the International Growth Inhibition Assay Reference Centre at the National Institutes of Health50,51. Synchronized Plasmodium falciparum 3D7 cultures at the late schizont stage were adjusted to 1.5% parasitemia, 4% hematocrit and 20 μl aliquots were added into 96 well flat bottom tissue culture plates. A Pfs48/45 specific humanized mAb TB31F52 was used as a negative control. Test antibody was added in triplicate wells over a concentration range from 1.0 to 0.0039 mg/ml (two-fold dilution series) and returned to culture (5% O2–5% CO2–90% N2 at 37 °C) for 40 h. Growth inhibition (parasitemia) was assessed by the lactate dehydrogenase activity assay53. The percent GIA was calculated using as: % GIA = 100–100 (sample A650 – uninfected RBC A650)/(infected control A650 – uninfected RBC A650).

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