DESCRIPTION

Used for the quantitative determination of Anti-MPXV E8L Human IgG concentration in serum and plasma.

PRINCIPLE OF THE ASSAY

This assay employs the quantitative indirect enzyme immunoassay technique. Recombinant MPXV E8L antigen has been pre-coated onto a microplate. Standards or samples are pipetted into the wells and any Anti-MPXV E8L Human IgG present is bound by the immobilized protein. After washing away any unbound substances, a biotin-labeled Mouse Anti-Human IgG antibody is added to the wells. After washing away any unbound substances, Streptavidin-HRP is added to the wells. Following a wash to remove any unbound enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of Anti-MPXV E8L Human IgG bound in the initial step. The color development is stopped and the intensity of the color is measured.

PRECISION

Intra-Assay Precision (Precision within an assay): <10% Three samples of known concentration were tested sixteen times on one plate to assess intra-assay precision. Inter-Assay Precision (Precision between assays): <15% Three samples of known concentration were tested in twenty four separate assays to assess inter-assay precision.

Sample		1	2	3	1	2	3
n		16	16	16	24	24	24
Mean (ng/mL)		9.9	2.4	1.2	9.7	2.4	1.1
Standard deviation		0.3	0.1	0.1	0.3	0.1	0.1
CV \(%\)		2.7	3.7	6.7	2.8	3.4	7.1

RECOVERY

The recovery of Anti-MPXV E8L Human IgG spiked to three different levels in human serum samples throughout the range of the assay was evaluated.

Sample	Dilution factor	Average (%)	Range (%)
Human serum (n=3)	1:1000	97	88-114
	1:2000	98	87-109

LINEARITY

To assess the linearity of the assay, human serum samples were spiked with high concentrations of Human Anti-MPXV E8L IgG and diluted with the Standard Diluent to produce samples with values within the dynamic range of the assay.

Sample	Dilution factor	Average (%)	Range (%)
Human serum (n=5)	1:2	102	95-115
	1:4	103	94-114
	1:8	102	91-108
	1:16	95	82-117