Table 3.

Binding ratio of four anti DBL1-VarO mAbs on a set of recombinant domains with differing length and mutated residues

Ref	Protein	PfEMP1-VarO Residues (No Cys)	Mutationsa	RBC bindingd	Fold increase Kd heparine	D15-50	E20-76	BD20E4	BDEE10	No tests
						Ratio to eDBL1f
[27]	eDBL1	2–487 (20)	#	+++	ND	1	1	1	1
[28]	eDBL1-s	2–471 (18)	# K87A	++	1	0.95	0.92	0.27	1.01	3
[22]	eDBL1-swt	1–471 (18)	Wild type	++++	ND	1	1	0.3	1	2
[22]	eHead	2–716 (31)	#	+++++	ND	1.4	1.5	1.3	1.8	2
						Ratio to parent eDBL1f
[22]	Mut K87	2–487 (20)	# K87A	++++	ND	0.93	0.85	0.90	0.87	1
[22]	Mut10fXab	2–487 (20)	# G66I Y67E V68G	Nil	ND	0.45	0.12	1.05	1.10	2
[22]	Mut11c	2–487 (20)	# G66I Y67E V68G	+++	ND	1.30	1.30	1.00	1.20	1
						Ratio to parent eDBL1-sf
[28]	Mut1	2–471 (18)	# K87A K2A0 K32A K40A	++	115	0.66	0.67	0.79	0.76	2
[28]	Mut2	2–471 (18)	# K87A K95A K166A K179A	Nil	1	0.47	0.36	1.05	1.1	3
[28]	Mut3	2–471 (18)	# K87A K423A K424A K451A K456A	++	218	0.94	0.91	1.1	0.85	2
[28]	Mut4	2–471 (18)	# K87A K226A K227A K230A	Nil	11	0.94	0.84	0.42	0.45	3
[28]	Mut5	2–471 (18)	# K87A K404A K407A K410A	++	1	0.85	0.77	1.03	0.96	2
[22]	Mut8	2–471 (18)	# K87A R64A Y67A R69A	Nil	ND	0.87	1.40	0.85	0.97	1
[22]	Mut9	2–471 (18)	# K87A T88A Y90A E92A	+	ND	1.03	1.05	0.90	1.00	1

ND not determined

^aPredicted N-glycosylation sites mutated (NxT/S to NxA)

^bMBP-Mut10 was cleaved by Factor Xa located at the cloning site and at the internal site introduced at positions 66–69 (IEGR)

^cDBL protein sequence identical to Mut10, but the cloning site (encoding a Factor Xa cleavage site) was changed for a thrombin cleavage site

^dFrom ref [22]

^eFrom ref [28]

^fThe mean ratio of reactivity was calculated as the mean of ratio to the parent protein observed for 3 successive dilutions in the linear part of the ELISA titration curves. The average ratio is shown for the proteins tested in 2-3 independent experiments