Development of a Novel Recombinant Full-Length IgY Monoclonal Antibody against Human Thymidine Kinase 1 for Automatic Chemiluminescence Analysis on a Sandwich Biotin-Streptavidin Platform for Early Tumour Discovery

Table 3

Comparative determination of the affinity binding and sensitivity (a) and specificity (b) between hTK1-IgY-pAb and the five different rmAbs (for the method, see Section 2.3.3).

(a) Comparison of sensitivity between hTK1-IgY-pAb (pAb) and the five hTK1-IgY-rmAbs (rmAb)

Type of IgY Ab	Affinity (rhTK1)		Sensitivity (calibrators)
	Kd value	Q^‡, <10^-9 Kd	Slope	Q^‡, <10%
pAb vs.		Yes	89.98
rmAb #1		Not	89.94	(0.04%) yes
rmAb #2		Not	83.51	(7.20%) yes
rmAb #3		Not	89.56	(0.46%) yes
rmAb #4		Yes	89.12	(0.95%) yes
rmAb #5		Yes	89.98	(0.00%) yes

(b) Comparison of specificity between hTK1-IgY-pAb (pAb) and the five hTK1-IgY rmAbs (rmAb) (TK1 in serum)

pAb vs. rmAb	value at low STK1p	Q^‡,	value at elevated STK1p	Q^‡,
rmAb #1	0.467	Not	0.512	Not
rmAb #2	0.181	Not	0.457	Not
rmAb #3	0.105	Not	0.409	Not
rmAb #4	0.964	Yes	0.890	Not
rmAb #5	0.920	Yes	0.963	Yes

The affinity binder had a Kd when high-affinity binding was achieved; sensitivity: when the slope (linearity of the calibrator curve) using the hTK1-IgY-rmAb declined less than 10%, it indicated that the rmAb was sufficiently sensitive; : value was tested by Pearson correlation. An value > 0.9 was considered a highly significant correlation. Kd: dissociation constant. ^‡Q: qualified.