Molecular dissection of the human antibody response to the structural repeat epitope of Plasmodium falciparum sporozoite from a protected donor

Table 2 V_H and V_L assignments and alignment of CDR 3 sequences The selected (NP 04, 12, 13 designated Pf NPNA-1 bind to the repeat epitope), all other NP clones were randomly picked after the panning procedure and were subsequently shown not to be reactive with the repeat epitope. Non-selected (R01-10) were randomly picked from the library prior to initiating panning. The peptide sequence of the heavy and light chain complementarity-determining region 3 (CDR3) is shown below. V_H/V_L families, segments and the number of differences from germline segments were determined by using the V BASE sequence directory (Tomlinson, I. M., Williams, S. C., Corbett, S. J., Cox, J. P. L. & Winter, G., MRC Centre for Protein Engineering, Cambridge, UK) and the DNAPLOT alignment package (Müller, W. & Althaus, H.-H., Köln University)

clone code*	V_H family	V_H Segment	Differences from germline	V_HCDR3	V_L family	V_L Segment	Differences from germline	V_LCDR3
PfNPNAl	VH3	DP46	10	DRDSSSYFDS	VkI	L12a	15	QQYNSYSGLT
NP04, NP12, NP13	VH3	DP46	10	DRDSSSYFDS	VkI	L12a		QQYNSYSGLT
R01	VH1	4M28^†‡	28(+6)* ^§-	DSESVAQWRY	VkIV	DPK24	43	QQSLSPVWT
R02	VH3	COS-3^‡	27 (+3)_	GVNWCSDY	VkI	DPK9	10	QQSYSTSWT
R03	VH5	DP73	35	LYTSIYYFDS	VkIV	DPK24	7	QQYYSTPLT
R04	VH3	DP46	8	DRVTNFWSGYFDY	VkIII	DPK22	13	QQYGSSPGFT
R05	VH3	DP58	23	DSTVKTVTKMRYGLD V	VkIII	DPK22	8	QQYGSSPFT
R06	VH1	4M28^†	12	DNYGDPGGGFDI	VkIII	DPK22	11	QQYGNSPRT
R07	VH5	DP73	9	RFWFGELYDAFDI	VkIV	DPK24	16	HQYYSTPQT
R08	VH5	DP73	34	LYTSIYYFDS	VkIII	DPK22	14	QQYGRSPWT
R09	VH3	V3-21^†	34	DQGGGWSSEVDS	VkIII	Vg	5	QQRSNWPLT
R10	VH1	DP7^‡	21 (+9)**	ALYGHDAFDI	VkI	DPK4	12	PKYNSALHT
NP02	VH3	DP47	36	ERPYDAFDS	VkIII	DPK22	23	QQYSTSPPMYN
NP03	VH5	DP73	40	LYTSIYYFDS	VkIII	Vg	17	KQRSKWPPIT
NP05	VH3	V3-48	14	EPRGAGTTLYFDY	VkIII	DPK22	22	QQYGGSPGYN
NP08	VH4	4.30^†	18	DRGVSSGWTFDC	VkII	DPK16	32	MQLTAFPWT
NP09	VH4	DP71	17	FRGGVAAGYDY	VkIII	DPK22	24	QHYRESCS
NP10	VH4	DP78	29	DRVRVPYYYIDV	VkIII	DPK22	15	QQYGTSPYS
NP11	VH3	VH3-8^†	12	DTTVTHYFDY	VkI	DPK9	21	QQSFSSPRT
NP14	VH1	DP88	20	GPGATIHYYYMDV	VkI	DPK8	18	QQLDNYPLT
NP15	VH5	DP73	36	LYTSIYYFDS	VkIII	DPK22	28	QQYGNSPPT

*Phage clones were either selected from the library at random (prefix R) or after four rounds of panning against (NPNA)₃C-BSA, eluting with free (NPNA)₃ peptide (prefix NP). ^† The segment given the best DNAPLOT match, although the segment sequence has not been verified by duplication. ^‡ Aligned after removal of the unusual sequence additions (see^§,-, ^‡,-, **). ^§ Figure in brackets indicates an unusual sequence addition. - Sequence has two additional codons in CDR1. - Sequence has one additional codon in CDR1. ** Sequence has three additional codons in CDR2.

ISSN: 1475-2875