Representative Human IL-22 ELISA Standard Curve from 0.78 - 50 pg/ml (see Technical Data Sheet)

Representative Standard Curves in Various Matrices (see Technical Data Sheet)

Percent Recovery of IL-22 in Various Matrices (see Technical Data Sheet)

Parallelism of Normal Serum (see Technical Data Sheet)

Disease Serum Parallelism (see Technical Data Sheet)

Linearity of Normal Human Serum (NHS) (see Technical Data Sheet)

Linearity of Disodium-EDTA Plasma (see Technical Data Sheet)

Linearity of Na-Heparin Plasma (see Technical Data Sheet)

Competitor Comparison of Representative Standard Curves (see Technical Data Sheet)

Competitor Comparison of Percent Recovery Curves (see Technical Data Sheet)

High detectability of endogenous IL-22 in patient serum samples (see Technical Data Sheet)

Human IL-22 ELISA, High Sensitivity (Serum, Plasma, TCM)

Catalog Number: 41701

Human Interleukin 22 ELISA designed for accurate quantitation of endogenous human IL-22 in healthy & disease sera/plasma, and tissue culture media samples with an LLOQ of 0.78 pg/ml. This assay measures unbound (free) IL-22. IL-22 Binding Protein (IL-22BP) may interfere with the assay if present at high concentrations.

Product Name: VeriKine-HS Human Interleukin 22 ELISA Kit

$655.00

Matrix Compatibility	Serum, Plasma, Tissue Culture Media
Assay Range	0.78 - 50 pg/ml
LLOQ	0.78 pg/ml Need more sensitivity? Check out our Sample Testing Services
Assay Length	4 hours
Specificity	Human IL-22

PBL’s IL-22 ELISA is intended for quantification of IL-22 in Tissue Culture Media (TCM), and complex matrices such as serum and plasma. The high-affinity form of the IL-22 binding protein (IL22-BP) does inhibit this assay which suggests the ELISA may measure free IL-22 in samples.

CVs and Spike Recovery	Inter-Assay: < 10% Intra-Assay: < 10% Spike recovery: > 96%
Cross-reactivity	Limited cross-reactivity with < 1% non-specific detection observed with monkey IL-22 (0.02% and 0.03% from two different runs) No cross-reactivity detected with Human IL-10, IL-17, IL-19, IL-20, IL-24, IL-26/AK155, IFN-β 1a, IFN-γ, IFN-ω Cynomolgus IFN-α 2 Cynomolgus/rhesus IFN-α Mouse IL-22, IFN-α A, IFN-β, IFN-γ Rat IL-22, IFN-α 1, IFN-α 14, IFN-β, IFN-γ Bovine IFN-Tau 2 or pig IFN-α.
Storage	2-8 °C
Expiration Date	One year from the date of manufacture
Shipping Condition	Wet Ice

Materials Provided

Pre-Coated Micro-Titer Stripwell Plate/Sealers
Wash Solution Concentrate
Human IL-22 Standard (10,000 pg/ml)
Standard Diluent
Assay Buffer
Antibody Concentrate
HRP Conjugate Concentrate
Concentrate Diluent
HRP Diluent
TMB Substrate Solution
Stop Solution

Additional Materials Required (Not Provided)

Microplate reader capable of reading an OD at a wavelength of 450 nm
Variable volume microtiter pipettes
Adjustable multichannel pipette (50-300 μl)
Reagent reservoirs
Wash bottle or plate washing system
Distilled or deionized water
Serological pipettes (1, 5, 10 or 25 ml)
Disposable pipette tips (polypropylene)
Plate shaker

Representative Standard Curve in Standard Diluent

Human IL-22 standard curves were prepared in Standard Diluent (SD) and assayed in triplicate over nine runs. The figure below shows the dose response and absorbance values. Error bars indicate the standard deviation.

Intra-Assay Precision
Sample	1	2	3	4
n	18	18	18	18
Mean (pg/ml)	26..68	6.42	2.86	4.52
Standard Deviation	1.38	0.62	0.16	0.18
CV (%)	5.2	9.7	5.5	4.1

Inter-Assay Precision
Sample	1	2	3	4	5
n	5	5	5	5	5
Mean (pg/ml)	28.05	6.73	3.13	4.80	5.79
Standard Deviation	3.62	0.68	0.17	0.27	0.54
CV (%)	12.9	10.1	5.3	5.7	9.3

Intermediate Precision
Sample	1	2	3	4	5
n	27	27	27	27	27
Mean (pg/ml)	31.93	7.76	3.43	5.30	6.45
Standard Deviation	4.34	1.24	0.24	0.62	0.94
CV (%)	13.6	16.0	12.3	11.8	14.6

Spike Recovery

TCM
Spike Sample	1	2	3
Target Conc. (pg/ml)	100	25	5
Mean Recovery	101.9	93.9	90.8
Range (%)	100.7-103.0	90.3-97.5	90.2-91.4
Disodium-EDTA Plasma
Spike Sample	1	2	3
Target Conc. (pg/ml)	100	25	5
Mean Recovery	97.3	88.7	83.9
Range (%)	87.6-102.6	82.0-93.3	76.1-92.5
Serum
Spike Sample	1	2	3
Target Conc. (pg/ml)	100	25	5
Mean Recovery	90.9	87.1	84.8
Range (%)	83.3-100.5	80.0-98.9	74.6-94.5

Representative Standard Curves in Various Matrices

Percent Recovery of IL-22 in Various Matrices

Parallelism of Normal Human Serum (NHS)

Three NHS samples with high levels of endogenous IL-22 were assayed in duplicate to assess reliable quantification after dilution within the standard curve range. Endogenous samples were diluted in Standard Diluent down to a 16-fold dilution. All samples fell within 100 ± 20% of the neat value. Error bars indicate the standard deviation.

Parallelism of Disease State Serum

Four Atopic Dermatitis (AD) serum samples with high levels of endogenous IL-22 were assayed in duplicate to assess reliable quantification after dilution within the standard curve range. Samples were diluted in Standard Diluent (SD) down to a 16-fold dilution. All samples fell within 100 ± 20% of the neat value. Error bars indicate the standard deviation.

Linearity of NHS

Linearity of Disodium-EDTA Plasma

Linearity of Na-Heparin Plasma

Representative Standard Curves of PBL and Competitor A

PBL’s and Competitor A’s standards were assayed on the 41701 PBL ELISA. Figure 13 shows the dose response curves and absorbance values in Standard Diluent (SD). Competitor A’s standard exhibits a lower dose response compared to PBL’s standard, as well as a lower percent recovery, which is less than 80% of the expected value. Competitor A’s standard curve demonstrated less sensitivity than PBL’s IL-22 ELISA.

Comparison of Percent Recovery Curves of PBL and Competitor A

Curves The figure below shows the recovery of the individual IL-22 standards at various concentrations. Competitor A’s standard was backfitted against PBL human cell-expressed standard, which was used to interpolate the concentrations of IL-22 and calculate the percent recovery of the curves from the expected value. PBL’s ELISA exhibited higher percent recovery than Competitor A.

PBL’s IL-22 ELISA (41701) Standard Curve in the Presence of IL22-BP

Competitor A’s Standard Curve in the Presence of IL22-BP

PBL’s IL-22 ELISA (41701) Recovery in the Presence of IL22-BP

Competitor A's Recovery in the Presence of IL22-BP

Healthy Donor Readability on PBL's ELISA
Sample Type	Mean of Quantified (pg/ml)	Range (pg/ml)	% Quantified	%Non-Quantified	% CV
NHS	1.84	0.80 - 5.43	96 (23/24)	4	5.6
Sodium-Citrate Plasma	2.54	1.06 - 24.60	100 (7/7)	0	4.5

Healthy Donor Readability on PBL's ELISA

Sample Type

Mean of Quantified (pg/ml)

Range (pg/ml)

% Quantified

%Non-Quantified

% CV

NHS

1.84

0.80 - 5.43

(23/24)

5.6

Sodium-Citrate Plasma

2.54

1.06 - 24.60

100

(7/7)

4.5

Healthy Donor Readability on Competitor A's ELISA
Sample Type	Mean of Quantified (pg/ml)	Range (pg/ml)	% Quantified	%Non-Quantified	% CV
NHS	20.43	16.26 - 24.60	8 (2/24)	92	2.8
Sodium-Citrate Plasma	ND	ND	0	100	ND

Healthy Donor Readability on Competitor A's ELISA

Sample Type

Mean of Quantified (pg/ml)

Range (pg/ml)

% Quantified

%Non-Quantified

% CV

NHS

20.43

16.26 - 24.60

(2/24)

2.8

Sodium-Citrate Plasma

100

ND: Not Detectable

Sample Type	Mean of Quantified (pg/ml)	Range (pg/ml)	% Quantified	%Non-Quantified	% CV
Disease State Readability on PBL's ELISA
AD	14.77	2.30 - 32.18	100	0	2.1
Psoriasis	5.33	1.25 - 25.99	100	0	3.8
RA	5.58	1.98 - 9.47	100	0	3.7
SLE	3.13	1.55 - 9.22	100	0	4.7

Sample Type	Mean of Quantified (pg/ml)	Range (pg/ml)	% Quantified	%Non-Quantified	% CV
Disease State Readability on Competitor A's ELISA
AD	32.42	14.28 - 73.07	60	40	5.0
Psoriasis	33.09	N/A	10	90	2.9
RA	ND	ND	0	100	ND
SLE	ND	ND	0	100	ND

Background

IL-22 is a member of the IL-10 family of cytokines¹. Other names include IL-TIF and Zcyto18. IL-22 is produced by a variety of cells including Th1, Th17, Th22 T-cells, NKT cells, ILC3, neutrophils and macrophages². Targeting primarily non-hemopoietic cells such as stromal and epithelial cells, IL-22 exerts both protective effects, such as hepatocyte and epithelial cell survival, and pro-inflammatory effects. It is also implicated in autoimmune diseases such as Systemic Lupus Erythematosus, Rheumatoid Arthritis and Psoriasis³. IL-22 is often considered a hallmark of IL-17 driven immune responses⁴.

References:

Dumoutier, L., et al. 2000. Genes & Immunity. 1(8): 488-94.

Dudakov J., Hanash A., van den Brink, M. 2015. Annual Review of Immunology. 33: 747-785.

Eyerich, K., Dimartino, V., Cavani, A. 2017. European Journal of Immunology. 47(4): 607-614.

Eyerich, S., et al. 2017. Allergologie Select. 1(1): 71-76.

Human IL-22 ELISA, High Sensitivity (Serum, Plasma, TCM)