Detection of cerebral spinal fluid‐associated chemokines in birth traumatized premature babies by chip‐based immunoaffinity CE

Abstract

A major concern in treating premature infants with birth-associated head trauma is the rapid determination of reliable biomarkers of neuroinflammation. To this end a chip-based immunoaffinity CE device has been applied to determine the concentrations of inflammation-associated chemokines in samples of cerebral spinal fluid collected from such subjects. The chip utilizes replaceable immunoaffinity disks, to which reactive antibody fragments (FAb) of 6 anti-chemokine-specific antibodies were immobilized. Following injection of a sample into the device, the analytes were captured by the immobilized FAbs, labelled in-situ with a red laser dye, chemically released and separated by CE. Each resolved peak was measured on-line by laser-induced fluorescence detection and the results compared to standard curves produced by running known chemokine standards through the immunoaffinity system. The complete processing of a sample took 10 min with separation of all 6 analytes being achieved in less than 2 min. The system compared well to commercial ELISA, analysis of the results by linear regression demonstrating r2 values in the range of 0.903 – 0.978 and intra- and inter-assay coefficients of variance (CV) of the migration times and the measured peak areas being less than 2.3% and 5%, respectively. Application of the system to analysis of cerebrospinal fluid from head traumatized babies clearly indicated the group with mild trauma versus those with severe injury. Additionally, CE analysis demonstrated that the severe trauma group could be divided into individuals with good and poor prognosis, which correlated with the clinical finding for each patient.

Developments in coupled solid‐phase extraction–capillary electrophoresis 2011–2013

“…An emerging trend is the development of chip‐based SPE–CE systems as the integration of SPE with microchip CE offers the advantages of improved sensitivity and sample clean‐up combined with relatively short analysis times . Nowadays, on‐chip labeling and purification of analytes can be performed in a fully automated way and the integration of such an approach with affinity extraction should yield an additional dimension of specificity, which may provide very effective methodologies for bioanalysis.…”

A decade of microchip electrophoresis for clinical diagnostics – A review of 2008–2017

“…Immune disorders including inflammatory diseases [95,96], allergic reactions and hypersensitivities [97][98][99][100][101], arthritis and gout [102,103] were investigated.…”

“…and in the cerebral spinal fluid of preterm babies with traumatic head injury[99]. The microfluidic chips combined affinity-based isolation of cytokines, fluorescence-labelling, and separation of labelled cytokines.…”

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2012–2014)

“…Phillips et al developed a chip‐based SPE‐CE system to isolate and determine six pro‐inflammatory chemokines from infants’ cerebrospinal fluids using reactive antibody fragments of antichemokine antibodies immobilized onto a 2‐mm glass fiber disk . The isolated analytes were labeled and eluted with acidic buffer to disrupt the complex releasing the chemokines for electrophoresis.…”