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Table 4 Expression levels and affinity constants assessed by SPR spectroscopy

From: Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity

  OcheA-9 OcheA-16 OcheA-17 OcheA-17B
Expression level (mg/kg FLM) 150 140 440 110
AMA1 DiCo1 KD (nM) 213 43
AMA1 DiCo2 KD (nM) 110 160 1750
AMA1 DiCo3 KD (nM) 68 55 61 1.7
Pf3D7 AMA1 KD (nM) 176
  1. To calculate expression levels the concentrations of recombinant antibodies OcheA-9, OcheA-16, OcheA-17 and OcheA-17B in N. benthamiana plant extracts were assessed measuring the capture level on a protein A surface in relation to an internal antibody standard using BiacoreT200. For affinity kinetic measurements purified recombinant antibodies OcheA, OcheA-9, OcheA-16, OcheA-17 and OcheA-17B were captured on a protein A surface (110, 110, 130, 140, 120 RU respectively) and the antigens were injected successively in concentrations of 360 nM, 180 nM and 90 nM for 180 s (30 µL/min). The surface was washed with HBS-EP buffer and regenerated with 30 mM HCl between measurements. Data were evaluated using T200EvaluationSoftware3.0 using the two state reaction binding model. “–”: not determinable