DESCRIPTION

CD30, also known as TNFRSF8 or Ki-1, is a transmembrane glycoprotein belonging to the tumor necrosis factor receptor superfamily. It is composed of an extracellular domain with six cysteine-rich repeats, a transmembrane segment, and an intracellular domain. CD30 is expressed on a small subset of activated T and B lymphocytes, and a variety of lymphoid neoplasms, with the highest expression in classical Hodgkin's lymphoma and anaplastic large cell lymphoma. The CD30 ligand (CD30L or CD153) is a type II transmembrane glycoprotein of the tumor necrosis factor ligand superfamily. Signal transduction of CD30/CD30L utilizes signal transducers, TNFR-associated factors (TRAF1, 2, 3 and 5).

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	1,075.9	30.0	2.8
2	8	256.6	8.5	3.3
3	8	140.2	9.4	6.7

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,071.7	75.8	7.1
2	16	258.0	12.7	4.9
3	16	137.6	7.7	5.6

RECOVERY

The recovery of human CD30/TNFRSF8 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:8	122	119-125
	1:16	121	117-126
Cell culture supernatant	1:4	122	115-130
	1:8	102	95-113

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Human plasma	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	115	103
	Range (%)	104-126	102-104
1:8	Average% of Expected	110	119
	Range (%)	105-115	108-130
1:16	Average% of Expected	105	106
	Range (%)	99-111	97-116

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human CD30/TNFRSF8 One-Step ELISA Kit produced at Sunlong Medical™.