SPARCL (Spatial Proximity Analyte Reagent Capture Luminescence) is a novel homogeneous proximity assay technology utilizing flash chemiluminescence detection without solid support or wash steps.

1. Lumigen SPARCL and Innova Biosciences
Lightning-Link®
Fast, Simple and Cost-Effective Immunoassay Development, Validation and Sample Analysis
SPARCL (Spatial Proximity Analyte Reagent Capture Luminescence) is a novel homogeneous proximity assay technology utilizing flash
chemiluminescence detection without solid support or wash steps. This format allows for rapid immunoassay development, in as little as 2
days. The homogenous format promotes fast antibody to antigen binding kinetics, allowing 30 minute incubation times. With the SPARCL
format, 8 or more assay development or validation exercises can be carried out in one day.
SPARCL is based on the interaction of a small molecule called acridan with horseradish peroxidase (HRP) in the presence of hydrogen
peroxidase. Acridan releases light energy proportional to the amount of analyte bound to the HRP and acridan labeled antibodies.
Lightning-Link from Innova Biosciences provides a simple, fast and easy kit that enables you to label your antibody or protein with HRP.
These world-leading Lightning-Link conjugation kits require only 30 seconds hands-on time. Furthermore, no clean-up of unbound HRP
is required, resulting in 100% antibody recovery. The conjugation of HRP to your antibody is complete and ready to use in 3 hours, allowing
assay development to start on the same day as labeling.
CASE STUDY: Rapid assay development for human CRP (C-Reactive Protein)
Day 1: Labeling 4 commercial antibodies with acridan and HRP using Lightning-Link , antibody screening and best pair selection.
Day 2: Titration of HRP-labeled and acridan conjugated antibodies, dilutional linearity testing.
Day 3: Establishment of dynamic ranges, selection of QC levels, MRD determination.
Day 4: Accuracy and Precision runs 1, 2 and 3, sample testing, parallelism testing.
Assay Characteristics:
Dynamic Range: 48 to 50,000 pg/mL (3 logs) means few sample repeat runs
MRD: 1:40
Sample Volume Requirement: 2 µl for dilution. 25 µl of diluted sample applied to assay
Assay Run Time: 30 minutes
Washing Steps: Zero
Plate Coating and Blocking: None
Equipment Required: 96- or 384-well plate reader, luminometer enabled, with injectors
22900 W. 8 Mile Road • Southfield MI 48033 USA • Telephone: 248 351-5600 or 877 LUMIGEN • Facsimile: 248 351-0518 • www.LUMIGEN.com
* Trademarks are property of their respective owners.

2. 22900 W. 8 Mile Road • Southfield MI 48033 USA • Telephone: 248 351-5600 or 877 LUMIGEN • Facsimile: 248 351-0518 • www.LUMIGEN.com
* Trademarks are property of their respective owners.
Figure 1. Human CRP SPARCL immunoassay. Note the standard curve has a range of 48.8 to 50,000 pg/mL.
CRP QC (pg/mL) 20,000 5000 200 Endogenous QC
Average 21196 5264 239 21678
%CV 6.8 8.7 13.7 5.3
RE 6 5.3 19.6 NA
Table 1. Accuracy and precision data for quality control samples (20,000, 5,000, 200 pg/ml and an endogenous sample) from 3 independent
assay runs. Concentration values were interpolated off the standard curve and individual concentration values were used to determine accuracy
(%CV) and precision (RE, Relative Error).