Test Results

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Test Results

  • Test Results
  • Compatibility
  • Batch to Batch

Comparison of ELISA Protocol

MicroFluere®

  • Use 20 µL or less
  • Fewer steps
MicroFluere® Incubation (min) Step
Capture 60 #1
Blocking 30 #2
Wash 0 #3
Standard 15 #4
Detection 15 #5
HRP 5 #6
Wash 0 #7
Wash 0 #8
Wash 0 #9
Substrate 5 #10
TOTAL 130 10
brackets1
brackets1
Capture antibody coating
90 minutes

Actual assay: 40 minutes

Traditional

  • Use 100 µL or more
  • More steps
Traditional Incubation* (min) Step
Capture 540 #1
Wash 0 #2
Wash 0 #3
Wash 0 #4
Blocking 60 #5
Wash 0 #6
Wash 0 #7
Wash 0 #8
Standard 120 #9
Wash 0 #10
Wash 0 #11
Wash 0 #12
Detection 120 #13
Wash 0 #14
Wash 0 #15
Wash 0 #16
HRP 20 #17
Wash 0 #18
Wash 0 #19
Wash 0 #20
Substrate 20 #21
TOTAL 880 21
brackets1
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Capture antibody coating 
Overnight + 60 minutes
Actual assay: 280 minutes

Summary Performance Data Comparison

Nb Manufacturer Kit Name Analyte Manufacturer Advertised Dynamic Range (pg/ml) Manufacturer Dynamic Range (pg/ml)
1 Thermo Fisher AAAV9 AAV9 affinity ligand 250-10000 62.5-8000, 125-16000
2 R&D Systems DY266 Human CXCL10/IP-10 31.2-2000 18.75-9600
3 R&D System DY202 Human IL-2 15.6-1000 18.7-9600
4 R&D Systems DY206 Human IL-6 9.6-600 9.6-9600
5 R&D System DY210 Human IL-8/CXCL8 31.2-2000 18.7-4800
6 R&D System DY208 Human TNF-alpha 15.6-1000 18.75-9600
7 R&D System DY293B Human VEGF 31.2-2000 37.5-1200
8 R&D System DY7360-05 HIV-1 Gag p24 15.6-1000 21.7-9322
9 R&D System DY3689 Rat TIM-1/KIM-1/HAVCR 15.6-1000 11.3-9000
10 R&D System DY564 Rat VEGF 15.6-1000 12.5-9000
11 R&D System DY247 Human IL-15 15.6-1000 21.7-9106
12 R&D System DY240 Human TGF-beta 1 31.2-2000 37.5-9600
13 R&D System DY212 Human GDNF 31.2-2000 75-9600
14 R&D System DY285 Human IFN-gamma 15.6-1000 37.5-9600
15 Sino Biologica SEKB10395 Human IL-6 2.34-150 1.56-1600
16 Sino Biological KIT11725A Human IFN-gamma 23.44-1500 23.44-12000
17 Sino Biological N/A SARS COV-19 N-Protein N/A 23.39-XXX
18 Sino Biological N/A SARS COV-19 S-Protein N/A 390-XXX
19 Sino Biological N/A SARS COV-19 IgG antibody N/A 1000-XXX

Substrate: QuantaRed™ Enhanced Chemifluorescent HRP Substrate Kit from Thermo Fisher

MicroFluere® Result Vs. Analytical Test Result of the Kit AAV9 Ligand Leakage

Plate by OptoFluidic

Normalized at lowest concentration

Plate by OptoFluidic

Normalized at highest concentration

MicroFluere® Kit’s Manual Note
Assay time 40 minutes 200 minutes excluding washing and handling time
LOD ≤0.125 ng/mL ≤1 ng/mL
Linear range 0.0625 – 4 ng/mL 0.25 – 4 ng/mL
Dynamic range 0.0625 – 8 ng/mL (MF 1000X Dec.)
0.125 – 16 ng/mL (MF 1250X Dec.)
0.25 – 16 ng/mL

Human CXCL10/IP-10 Quantikine ELISA Kit

Plate by OptoFluidic
31.2-2000 pg/mL
Plate by OptoFluidic
  • R-square: 0.99823
  • Dynamic range: 75-9,600 pg/mL
  • Sample/reagent volume: ≤20µL
  • Assay time: 40 minutes

Human IL-2 DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99844
  • Dynamic range: 18.7-9,600 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human IL-6 DuoSet ELISA

Plate by OptoFluidic

9.4-600 pg/mL

Plate by OptoFluidic

Human IL-8/CXCL8 DuoSet ELISA

Plate by OptoFluidic

31.2-2000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99941
  • Dynamic range: 18.75-4,800 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human TNF-alpha DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99867
  • Dynamic range: 18.75-9,600 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes
Plate by OptoFluidic
  • R-square: 0.99867
  • Dynamic range: 18.75-4,800 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human VEGF DuoSet ELISA

Plate by OptoFluidic

31.2-2000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99867
  • LOD: 12.5 pg/mL
  • Dynamic range: 12.5-9,000 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

HIV-1 Gag p24 DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99986
  • LOD: 21.1 pg/mL
  • Dynamic range: 21.7-9,322 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Rat TIM-1/KIM-1/HAVCR DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99081
  • LOD: 11.3 pg/mL
  • Dynamic range: 11.3-9,000 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Rat VEGF DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99784
  • Dynamic range: 37.5-1200 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human IL-15 DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99838
  • LOD: 21.7 pg/mL
  • Dynamic range: 21.7-9,106 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human TGF-beta 1 DuoSet ELISA

Plate by OptoFluidic

31.2-2000 pg/mL

Plate by OptoFluidic
  • R-square: 0.99529
  • Dynamic range: 37.5-9600 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human GDNF DuoSet ELISA

31.2-2000 pg/mL

Plate by OptoFluidic
  • R-square: 0.97313
  • Dynamic range: 75-9,600 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Human IFN-gamma DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Plate by OptoFluidic
  • R-square: 0.9976
  • Dynamic range: 37.5-9,600 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

MicroFluere® Vs. SEKB10395 kit – Human IL-6

Conc. (pg/mL) Zero standard subtracted (FL)
0 0
1.5625 121
3.125 200
6.25 485
12.5 1103
25 1912
50 3790
100 7753
200 15599
400 29655
800 53358
1600 91553
3200 133184
6400 146631
Plate by OptoFluidic
Plate by OptoFluidic
Concentration (pg/mL) Zero standard subtracted OD
0 0
2.34 0.033
4.69 0.072
9.38 0.154
18.75 0.310
37.5 0.612
75 1.184
150 2.113
 MicroFluere®
SEKB10395
Note
Assay time35 minutes200 minutesexcluding washing and handling time
LOD1.93 pg/mL2.34 pg/mLZero standard + 3β
Linear range1.56 – 1600 pg/mL2.34 – 150 pg/mL

HRP -conjugated detection antibody – Human IFNg

Conc. (pg/mL)

A

B

C

Avg

STD

CV%

0

1723

1352

1371

1482

208.9282

14%

5.85938

1841

1613

1573

1675.667

144.5729

9%

11.71875

2254

2024

1690

1989.333

283.5936

14%

23.4375

2831

2094

2458

2461

368.5092

15%

46.875

3209

2936

2489

2878

363.4873

13%

93.75

4617

4293

3677

4195.667

477.499

11%

187.5

7180

6929

7280

7129.667

180.8323

3%

375

11235

11274

9543

10684

988.3274

9%

750

18098

17855

15684

17212.33

1329.14

8%

1500

34187

35273

31240

33566.67

2086.835

6%

3000

64935

61310

58076

61440.33

3431.357

6%

6000

104089

103277

96657

101341

4076.73

4%

12000

131987

137376

123600

130987.7

6942.157

5%

24000

148590

145788

136413

143597

6377.319

4%

Plate by OptoFluidic
Plate by OptoFluidic
MicroFluere® KIT11725A Note
Assay time 35 minutes 200 minutes excluding washing and handling time
LOD 6.9 pg/mL 15.1 pg/mL Zero standard + 3β
Linear range (4PL range) 23.44 – 6000 pg/mL (23.44-12000 pg/mL) 23.44 – 1500 pg/mL (NA)

COVID-19 N-protein ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

061920
LOD: 0.02839 ng/mL
R2: 0.99785
COGS of reagent: $25/plate

Note: The reagent cost are calculated based on retail price.

COVID-19 S-protein ELISA

Plate by OptoFluidic
  • R-square: 0.99794
  • Dynamic range: 390-XXX pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Mouse TGF-β1 ELISA Experiment at Prof. Tae-Hwa Chun’s Lab

Plate by OptoFluidic

31.2 – 2,000 pg/mL

Plate by OptoFluidic

Dynamic range = ~50 pg/mL – 15,000 pg

Human IFN-alpha 2/IFNA2 DuoSet ELISA

Plate by OptoFluidic
3.1-200 pg/mL

Human IL-10 DuoSet ELISA

Plate by OptoFluidic

31.2-2000 pg/mL

Human TGF-beta 1 DuoSet ELISA

Plate by OptoFluidic

15.6-1000 pg/mL

Compatibility tested commercial ELISA readers

  1. EnSpire 2300 Multimode Plate Reader (PerkinElmer)
  2. Infinite F200 Fluorescent Microplate Reader System (Tecan)
  3. Synergy Neo2 Hybrid Multi-Mode Reader (BioTek Instruments)
  4. Synergy H1 Hybrid Multi-Mode Reader (BioTek Instruments)
  5. Synergy HT (BioTek Instruments)
  6. GloMax®-Multi+ Detection System (Promega)
  7. Varioskan Flash (Thermo Fisher Scientific)

Intra- assay CVs

 

Plate 1

Conc. (pg/mL)

Rep 1

Rep 2

Rep 3

Avg

STD

CV%

0

399

365

412

392

24

6.2%

3.125

511

464

464

480

27

5.7%

6.25

585

550

461

532

64

12.0%

12.5

661

667

704

677

23

3.4%

25

923

1027

911

954

64

6.7%

50

1452

1510

1536

1499

43

2.9%

100

2287

2619

2607

2504

188

7.5%

200

4638

4672

4234

4515

244

5.4%

Plate1

Plate 2

Conc. (pg/mL)

Rep A

Rep B

Rep C

Avg

STD

CV%

0

450

379

369

399

44

11.1%

3.125

454

439

401

431

27

6.3%

6.25

592

471

474

512

69

13.5%

12.5

763

587

680

677

88

13.0%

25

1008

993

872

958

75

7.8%

50

1710

1581

1514

1602

100

6.2%

100

2957

2334

2665

2652

312

11.8%

200

4702

4807

5020

4843

162

3.3%

Plate2

Inter- assay CVs

Conc. (pg/mL)

Plate 1

Plate 2

Avg

STD

CV%

Rep 1

Rep 2

Rep 3

Rep A

Rep B

Rep C

0

399

365

412

450

379

369

396

32

8.1%

3.125

511

464

464

454

439

401

456

36

7.9%

6.25

585

550

461

592

471

474

522

60

11.6%

12.5

661

667

704

763

587

680

677

58

8.5%

25

923

1027

911

1008

993

872

956

62

6.5%

50

1452

1510

1536

1710

1581

1514

1551

89

5.7%

100

2287

2619

2607

2957

2334

2665

2578

244

9.5%

200

4638

4672

4234

4702

4807

5020

4679

258

5.5%

Plate1-2
Plate1and2