Construction of human liver cancer vascular endothelium cDNA expression library and screening of the endothelium-associated antigen genes

Table 1 Titer of anti-HLCVECs antibody in immunized animals

	D21	D28	D35	D42	D49	D56
HLCVEC	1:1000	1:3000	1:9000-	1:9000	1:27000	1:27000+

1:9000- indicates that the titer is lower than 1:9000, 1:27000+ indicates that the titer is 1:27000 or higher.

Table 2 Construction of HLCVECs libraries in λ-ZAP expression vector and the number of SEREX-identified antigens

Patient	Age (years)	Primary size	Screening serum	Clones screened	Positive clones	cDNA fragment	Known genes	Unknown genes
HCC	56	2 × 106	immunized	6 × 105	36	18	15	3

Table 3 Results of immunoscreening of HLCVECs cDNA expression library by immunized BALB/C sera screening

Clone	Size (bp)	Gene	Localization	Identity	Accession number
EC19	2100	HS.Protein disulfide	7q35	99%	NM 004911.2
		isomerase related protein
EC24	700	HS.Glucose regulated protein	15q15	98%	NM 0O5313.3
EC26	1000	HS.Chemokine ligand 1	4q12-q13	99%	BC 011976
EC29	1600	HS.Inner membrane protein	2p11.2	99%	NM 006839.1
EC30	1800	HS.Hypothetical protein Loc283241	11q12.3	99%	XM 208579.1
EC31	1700	HS.genomic DNA, clone	11q	98%	AP 002340.3
EC35	700	HS.BMP-6	15q13-q15	98%	NM 013372.1
EC36	2200	HS.Glyceral dehyde-3-	12p13.31	98%	BC 004109
		phosphate dehydrogenase
EC38	1900	HS.Replication factor C5	12q24.2	99%	NM 007370.2
EC39	2100	HS.BAC, CloneRP11-1591120		99%	AC 010974.9
EC40	800	HS.Sjogren syndrome antigen B	2q31.1	98%	BC 020818
EC42	2400	HS.X-ray repair complementing	12q23	99%	AY 034001.1
		defective repair in Chinese
EC51	400	HS.S100 calcium-binding protein A6	1q21	100%	BC 009017
EC52	1500	HS.Protein kinase C	19p1.1	98%	NM 002743.1
EC53	1700	HS.Heat shock 70 ku protein4	5q31.3	99%	XM 114482.2
EC59	1900	HS.DVS27 related protein	9p24.1	99%	BC 04785.1
EC62	1500	HS.NAC alpha mRNA	12q2	99%	AY 034001.1
EC63	1500	HS.Hypothetical protein DJ1042k10.2	2213.1	100%	HS 1042k10