Mimix™ EGFR Multiplex cfDNA Set Reference Standard

The EGFR Multiplex cfDNA Reference Standard Set is a cell line-derived, clinically relevant control that can be used to assess the performance of cfDNA assays that detect somatic resistance mutations in EGFR.

High-throughput platforms such as PCR-based assays and Next Generation Sequencing (NGS) give laboratories much greater power when profiling cfDNA samples. However, small quantities of DNA in combination with complex variants at low allelic frequencies represents one of the major challenges users encounter when profiling cfDNA. Horizon Discovery has developed a comprehensive cfDNA Reference Standard range to support the development and continued validation of cfDNA assays. Our cfDNA products are derived from human cell lines and fragmented to an average size of 160 bp to closely resemble cfDNA extracted from human plasma. The EGFR Multiplex cfDNA Reference Standard is supplied at 5%, 1%, 0.1% and 0% (EGFR Multiplex wild type) allelic frequencies and covers ten EGFR variants implicated in the responsiveness to EGFR tyrosine kinase inhibitors (EGFR-TKIs) and anti-EGFR monoclonal antibodies. This product covers clinically relevant SNPs, insertions and deletions in EGFR and can be used to optimize, validate or routinely monitor assay performance.

With this product, you are able to:

Analyze the sensitivity and specificity of your assay
Gain certainty of the limit of detection and limit of quantification
Optimize and validate new EGFR-specific assays and routinely monitor their performance

Technical Data

Format: cfDNA

Genes Covered: EGFR

Allelic Frequencies: 5%, 1%, 0.1% and 0% (EGFR Multiplex wild type)

Buffer: Tris-EDTA (10mM Tris-HCl, 1mM EDTA), pH 8.0

Product Information

Fragment Size: 160 bp

Unit Size: 350 ng per vial

Concentration: 20 ng/µl

Verified Mutations:

Chromosome	Gene	Variant	Expected Allelic Frequency
EGFR Multiplex wild type cfDNA Reference Standard: HD824
7p12	EGFR	L858R	0.0%
7p12	EGFR	ΔE746 - A750	0.0%
7p12	EGFR	T790M	0.0%
7p12	EGFR	V769 - D770insASV	0.0%
7p12	EGFR	L861Q	0.0%
7p12	EGFR	G719S	0.0%
7p12	EGFR	C797S	0.0%
7p12	EGFR	S464L	0.0%
7p12	EGFR	G465R	0.0%
7p12	EGFR	S768I	0.0%

Chromosome	Gene	Variant	Expected Allelic Frequency
5% EGFR Multiplex cfDNA Reference Standard: HD821
7p12	EGFR	L858R	5.0%
7p12	EGFR	ΔE746 - A750	5.0%
7p12	EGFR	T790M	5.0%
7p12	EGFR	V769 - D770insASV	5.0%
7p12	EGFR	L861Q	5.0%
7p12	EGFR	G719S	5.0%
7p12	EGFR	C797S	5.0%
7p12	EGFR	S464L	5.0%
7p12	EGFR	G465R	5.0%
7p12	EGFR	S768I	5.0%

Chromosome	Gene	Variant	Expected Allelic Frequency
1% EGFR Multiplex cfDNA Reference Standard: HD822
7p12	EGFR	L858R	1.0%
7p12	EGFR	ΔE746 - A750	1.0%
7p12	EGFR	T790M	1.0%
7p12	EGFR	V769 - D770insASV	1.0%
7p12	EGFR	L861Q	1.0%
7p12	EGFR	G719S	1.0%
7p12	EGFR	C797S	1.0%
7p12	EGFR	S464L	1.0%
7p12	EGFR	G465R	1.0%
7p12	EGFR	S768I	1.0%

Chromosome	Gene	Variant	Expected Allelic Frequency
0.1% EGFR Multiplex cfDNA Reference Standard: HD823
7p12	EGFR	L858R	0.1%
7p12	EGFR	ΔE746 - A750	0.1%
7p12	EGFR	T790M	0.1%
7p12	EGFR	V769 - D770insASV	0.1%
7p12	EGFR	L861Q	0.1%
7p12	EGFR	G719S	0.1%
7p12	EGFR	C797S	0.1%
7p12	EGFR	S464L	0.1%
7p12	EGFR	G465R	0.1%
7p12	EGFR	S768I	0.1%

An additional EGFR mutation is detectable in this reference standard. The allelic frequency of the mutation is not reported on the Certificate of Analysis. We include the details here to allow our customers to assess any impact on their work flows when using this product.

AA Mutation	CDS mutation	GRCh67 Location	rsID	Cause
Q787Q	c.2361G>A	7:55249063	rs1050171	Endogenous variant

General Information

Storage: 4°C

Expiry: See all product shelf life information

Quality Control

Fragmentation Size: D1000 DNA Screen Tape assay

Allelic Frequency: Droplet Digital™ PCR

Quantification: Qubit dsDNA BR Assay (post-fragmentation)

Intended use: For assay developers and molecular diagnostic labs for routine performance monitoring of molecular biology assays. Research use only. Not for diagnostic procedures.