Functional recombinant MHC class II molecules and high-throughput peptide-binding assays

Justesen, Sune; Harndahl, Mikkel; Lamberth, Kasper; Nielsen, Lise-Lotte B; Buus, Søren

doi:10.1186/1745-7580-5-2

Immunome Research

Table 1 Affinity measurements using LOCI assay.

From: Functional recombinant MHC class II molecules and high-throughput peptide-binding assays

		DR1GrpE^a Komp^b	DR2a Komp^b	DR2a pH6	DR3	DR4	DRB1*0813	DRB3*0301	DQA10501 DQB10201	DP401	I-E^D	Reference
Designation	Sequence	IC50 (nM)	IC50 (nM)	nM	nM	nM	nM	nM	nM	nM	nM
MBP	FLYGALLLA	3921	NB	NB	NB	NB	NB	N.D	N.D	34	NB	[26]
TERT572Y	YLFFYRKSV	4287	NB	2648	NB	1593	95	N.D	N.D	NB	322
Flu PR8 derived	FMYSDFHFI	642	1018	637	NB	1130	NB	N.D	N.D	1	NB	[27]
Pox.o42.con A2	FLIDLAFLI	NB	4276	NB	NB	NB	NB	NB	NB	179	NB
Pox.o42.con.im. A2	RMIAISAKV	1464	1985	1647	516	NB	267	1259	NB	NB	NB
Brucella	WMARRHAER	NB	NB	NB	NB	NB	224	NB	NB	NB	6
Salmonella	MMAWRMMRY	7	25	18	1709	68	28	1426	4318	NB	837
Flu HA1194-202	IVFMWAIHH	42	529	167	NB	NB	276	NB	NB	NB	NB
Hantaan	FMSRKLHRY	NB	2552	NB	279	NB	21		NB	NB	3
Oxytocinase271	YEKKYFAATQFEPLAARL	5	29	6	38	6	38	33	1090	1	NB	[79] (DP401)
Thyroid peroxidase	YIDVWLGGLAENFLPY	605	691	2138	NB	NB	NB	3184	564	602	NB	[6]
No nat. source	YPFIEQEGPEFFDQE	1279	NB	NB	289	3972	NB	3228	648	NB	NB	[6]
Hepatitis B	YLCQVFADATPTGWGL	1850	NB	3703	69	18	2988	2058	1335	13	NB	[6] (DR3, DR4)
Plasmodium Falciparum	YILLKKILSSRFNQM	7	2	4	32	94	4	1	NB	1	20	[28](DR1, DR2a, DR4)
No natural source	YILFLVKMNALRRLPV	10	20	5	39	?	78	3	NB	45	582	[28] (DR1, DR2a, DR4)
Hepatitis B	VGNFTGLYSSTVPVF	3	31	102	1414	632	2892	9	635	NB	NB
Influenza	PLKAEIAQRLEDV	50	41	65	134	2678	NB	11	NB	2091	NB	[29] (DR1, DR2a)
HIV	PIVQNIQGQMVHQ	71	852	447	471	NB	NB	23	NB	NB	NB	[30](DR1)
HepatitisB	PDRVHFASPLHVAWR	5	26	8	6	881	39	3	2807	124	NB
Clostridium tetanus	QYIKANSKFIGITE	18	11	22	30	432	29	1	NB	56	1501	[29] (DR1, DR2a)
No nat. source	KILEPFRKYTAFTIP	10	1	16	18	1600	11	1015	1542	30	2198	[34] (DR2a)
Hepatitis B	AFSYMDDVVLGAKSV	215	3382	1721	87	26	2753	764	2039	NB	NB
Mycobact leprea	GVTYEIDLTNKN	NB	NB	NB	4346	2548	NB	NB	NB	NB	NB	[6] (DR4)
Murine β₂m	IQMLKNGKKIPKVEMS	1232	3	6	21	NB	73	70	NB	NB	117	[31] (I-Ed)
Lambda repressor	YLEDARRLKAIYEKKK	2999	34	87	1098	3120	136	NB	NB	NB	184	[31] (I-Ed)
6xH Lambda 2(315)	HHHHHHIEGRFAALWFRNHFVFGGGTK	4	20	5	882	11	8	1198	NB	2	26	[32]
6xH Influenza A (PR8) HA 109–119	YHHHHHHIEGRSFERFEIFPKE	325	23	9	1429	1869	181	1528	3913	1	984	[33]
CLIP 6xH	LPKPPKPVSKMRMATPLLMQALPMYHHHHHH	1	1	1	1	1	2	1	224	1	23
CLIP	YLPKPPKPVSKMRMATPLLMQALPM	1	1	1	1	2	2	1	381	4	72
HA306-318	YKYVKQNTLKLAT	7	2	10	13	8	11	4	NB	NB	25	Most DR

Values represent an average of 2–4 measurements. Affinity measurements higher than 5000 nM were categorized as non-binders (NB). N.D: Not determined. a: The DR1 allele used for these measurements was produced as a construct comprising amino acids 1–181 and 1–190 of α and β chain respectively fused C terminally to the dimerizing nucleotide exchange factor GrpE (see additional file 1), this construct was not biotinylated and used in the LOCI competition assay. b: Determined using competitive LOCI assay. The reference column also states the MHC alleles known to bind the peptide.

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