TY - JOUR
T1 - A quantitative lateral flow assay to detect complement activation in blood
AU - Schramm, Elizabeth C.
AU - Staten, Nick R.
AU - Zhang, Zhouning
AU - Bruce, Samuel S.
AU - Kellner, Christopher
AU - Atkinson, John P.
AU - Kyttaris, Vasileios C.
AU - Tsokos, George C.
AU - Petri, Michelle
AU - Sander Connolly, E.
AU - Olson, Paul K.
N1 - Funding Information:
The authors thank Steven Wagner and Michael Takes for technical assistance. This work was supported in part by a grant from the National Institutes of Health ( HL103378 to P.K.O.).
Publisher Copyright:
© 2015 Elsevier Inc. All rights reserved.
PY - 2015/5/15
Y1 - 2015/5/15
N2 - Complement is a major effector arm of the innate immune system that responds rapidly to pathogens or altered self. The central protein of the system, C3, participates in an amplification loop that can lead to rapid complement deposition on a target and, if excessive, can result in host tissue damage. Currently, complement activation is routinely monitored by assessing total C3 levels, which is an indirect and relatively insensitive method. An alternative approach would be to measure downstream C3 activation products such as C3a and iC3b. However, in vitro activation can produce falsely elevated levels of these biomarkers. To circumvent this issue, a lateral flow immunoassay system was developed that measures iC3b in whole blood, plasma, and serum and avoids in vitro activation by minimizing sample handling. This assay system returns results within 15 min and specifically measures iC3b while having minimal cross-reactivity to other C3 split products. While evaluating the potential of this assay, it was observed that circulating iC3b levels can distinguish healthy individuals from those with complement activation-associated diseases. This tool is engineered to provide an improved method to assess complement activation at point of care and could facilitate studies to monitor disease progression in a variety of inflammatory conditions.
AB - Complement is a major effector arm of the innate immune system that responds rapidly to pathogens or altered self. The central protein of the system, C3, participates in an amplification loop that can lead to rapid complement deposition on a target and, if excessive, can result in host tissue damage. Currently, complement activation is routinely monitored by assessing total C3 levels, which is an indirect and relatively insensitive method. An alternative approach would be to measure downstream C3 activation products such as C3a and iC3b. However, in vitro activation can produce falsely elevated levels of these biomarkers. To circumvent this issue, a lateral flow immunoassay system was developed that measures iC3b in whole blood, plasma, and serum and avoids in vitro activation by minimizing sample handling. This assay system returns results within 15 min and specifically measures iC3b while having minimal cross-reactivity to other C3 split products. While evaluating the potential of this assay, it was observed that circulating iC3b levels can distinguish healthy individuals from those with complement activation-associated diseases. This tool is engineered to provide an improved method to assess complement activation at point of care and could facilitate studies to monitor disease progression in a variety of inflammatory conditions.
KW - Complement activation
KW - Intracerebral hemorrhage (ICH)
KW - Lateral flow assay
KW - Lupus
KW - iC3b biomarker
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U2 - 10.1016/j.ab.2015.01.024
DO - 10.1016/j.ab.2015.01.024
M3 - Article
C2 - 25660530
AN - SCOPUS:84961340409
SN - 0003-2697
VL - 477
SP - 78
EP - 85
JO - Analytical biochemistry
JF - Analytical biochemistry
ER -