HBeAg gene expression with baculovirus vector in silk worm cells

Table 1 HBeAg antigenicity detection with ELISA

Sample	Dilution
	1:1000	1:2000	1:4000	1:8000	1:16000	1:32000	1:64000
Cell culture medium	+	+	+	+	+	+	-
Culture cell lysate	+	+	-	-	-	-	-

Table 2 Comparison of two methods in anti-HBe antibody detection

Conventional method	n	Positive (n)
		Double sandwich methoda	Indirectb
Anti-HBe(+) and anti-HBc(-)	35	35	35
Anti-HBe(-) and anti-HBc(+)	52	4	52
Anti-HBe(+) and anti-HBc(+)	37	37	37
Anti-HBe(-) and anti-HBc(-)	42	2	0

^aThe purified HBeAg expressed in karyotic cells was used to coat enzyme-labeled reaction plate (100 ng/well), incubated throughout the night at 4 °C, and then serum to be detected was added to it. After 30 min at 40 °C, HRP-Labeled HBeAg expressed in karyotic cells was added to it, and at last this was colorized by application of TMB-H₂O₂. Those P/N ≥ 2.1 were positive.

^bThe anti-HBe antibody detection kit for sale in market employed the competition inhibition method.

Table 3 Purification of HBeAg Produced by Bm N cells

Method	Total activity (by ELISA)	Specific activity (per mg protein)	Purification factor		Total recovery (%)
			A	B
Cell culture medium	1:1.60 × 106	1:2.9 × 105	1.0	/	/
(NH4)2SO4	1:1.32 × 106	1:8.5 × 105	2.9	2.9	80
Sephacryl S-200	1:1.12 × 106	1:5.3 × 104	6.3	18.3	70
DEAE Sepharose FF	1:8.40 × 105	1:5.0 × 105	9.4	172.0	52

A: Individual steps; B: Accumulated results.

Table 4 Characteristics of HBeAg expressed in E. coli cells as compared with those in karyotic cells with the double-antibody Sandwich

Antigen	Cell	HBeAg kit	Anti-HBc kit
HBeAg 1	karyotic	10-7	10-1
HBeAg 2	E. coli	10-5	10-3