A simplified immunoprecipitation method for quantitatively measuring antibody responses in clinical sera samples by using mammalian-produced Renillaluciferase-antigen fusion proteins

Table 1 Immunoprecipitation capacity of 1 μl of human sera for Ruc-tumor antigen fusion proteins^a

		Ruc	p53	K-Ras	Smad4	β-CAT-Δ1	c-Myc
Controls	1	194	19,319	480	10,582	269	4,752
	2	9	9,830	1,064	3,575	835	2,913
	3	8	5,236	445	1,773	211	2,006
	4	38	3,187	477	1,919	530	1,831
	5	14	11,908	795	6,884	161	3,346
	6	31	5,390	823	1,724	235	2,050
	7	76	22,526	1,909	6,996	259	11,816
	8	29	15,338	943	8,043	445	3,475
	9	10	12,282	1,162	19,380	215	3,623
	10	9	11,130	1,109	4,429	501	5,060
+ 3 SD^b		214	30,234	2,237	22,788	997	12,874
Head and Neck	11	0	10,904	508	2,721	196	2,193
	12	0	31,593 ^c	738	4,822	465	3,801
	13	0	12,367	840	1,868	673	4,407
	14	13	14,705	1,012	5,666	195	1,837
	15	33	31,733 ^c	1,189	5,264	552	4,107
	16	121	4,828	621	980	279	1,974
	17	0	8,517	1,160	8,396	336	2,958
	18	0	19,240	1,283	9,485	327	1,814
	19	0	11,224	1,517	4,454	410	4,370
	20	28	7,322	554	2,261	723	2,343
Breast	21	44	13,211	960	10,219	308	5,988
	22	10	18,814	696	42,970 ^c	302	5,450
	23	38	14,598	608	8,484	339	4,336
	24	77	11,587	1,655	17,297	2363 ^c	3,431
	25	17	19,954	532	10,184	772	15,650 ^c
	26	25	9,538	195	5,962	300	1,646
	27	10	7,815	2,561 ^c	20,628	426	3,524
	28	21	15,607	308	7,380	284	1,579
	29	0	18,058	160	6,790	304	2,333
	30	245 ^c	25,479	1,919	9,727	495	3,787
Colon	31	4	6,656	1,204	3,252	267	1,763
	32	40	20,928	4,293 ^c	5,567	962	6,143
	33	42	34,703 ^c	1,472	10,830	716	4,906
	34	51	300,943 ^c	6,439 ^c	2,610	992	3,789
	35	35	5,670	3,306 ^c	3,860	477	1,772
	36	44	6,516	695	37,344 ^c	371	2,395

^aSera, FLAG-Ruc-fusion extracts, protein A/G beads and buffer were mixed together, incubated for 60 minutes and processed. The data, light units, is the average of two experiments and is corrected for background (beads plus extract, but no sera). The standard deviation for each value is also available (see Additional file 1).
^bValues of the averages of the 10 control sera plus 3 standard deviations.
^cNumbers in bold are statistically significant: greater than the average plus 3 standard deviations of the 10 control sera.

ISSN: 1472-6750