ac9b02572_si_001.pdf (1006.82 kB)
Centrifugation-Assisted Immiscible Fluid Filtration for Dual-Bioanalyte Extraction
journal contribution
posted on 2019-08-24, 14:29 authored by Duane
S. Juang, Scott M. Berry, Chao Li, Joshua M. Lang, David J. BeebeThe
extraction of bioanalytes is the first step in many diagnostic
and analytical assays. However, most bioanalyte extraction methods
require extensive dilution-based washing processes that are not only
time-consuming and laborious but can also result in significant sample
loss, limiting their applications in rare sample analyses. Here, we
present a method that enables the efficient extraction of multiple
different bioanalytes from rare samples (down to 10 cells) without
washingcentrifugation-assisted immiscible fluid filtration
(CIFF). CIFF utilizes centrifugal force to drive the movement of analyte-bound
glass microbeads from an aqueous sample into an immiscible hydrophobic
solution to perform an efficient, simple, and nondilutive extraction.
The method can be performed using conventional polymerase chain reaction
(PCR) tubes with no requirement of specialized devices, columns, or
instruments, making it broadly accessible and cost-effective. The
CIFF process can effectively remove approximately 99.5% of the aqueous
sample in one extraction with only 0.5% residual carryover, whereas
a traditional “spin-down and aspirate” operation results
in a higher 3.6% carryover. Another unique aspect of CIFF is its ability
to perform two different solid-phase bioanalytes extractions simultaneously
within a single vessel without fractionating the sample or performing
serial extractions. Here we demonstrate efficient mRNA and DNA extraction
from low-input samples (down to 10 cells) with slightly higher to
comparable recovery compared to a traditional column-based extraction
technique and the simultaneous extraction of two different proteins
in the same tube using CIFF.