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Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit

Designed for high efficiency cfDNA isolation 

For research use only. Not for use in diagnostic procedures.

Performance Data

The superb performance of Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit has been extensively validated.  Please review the examples of publications and performance data below.  For a full list of publications, please visit the Publications page.

Performance Assessment

Apostle MiniMax technology offers a best-in-class efficiency and purity compared with conventional technologies to capture and isolate the circulating cell-free genetic materials. It has been trusted by many of the world's most prestigious leaders in life sciences.

"...This includes recognizing that an innovative product from Apostle is the best technology to address these needs." said Steve Wowk, Director of Genomics, Beckman Coulter Life Sciences.


Some Examples of Published Performance Assessment Data


(Figure 4) - "Most notably, the Apostle particles outperformed all others, achieving almost 2-fold higher recovery yields than the particles supplied in the X kit. "

This link directs to an external page, reviewing the performance and data of Apostle MiniMax by Beckman Coulter Life Sciences.

(Figure 1) "For each tube the Apostle MiniMax extracted higher total yield of DNA. "


This link directs to an external Application Note. This application note compares workflows and yield for the extraction of cfDNA using Apostle MiniMax High Efficiency cfDNA Isolation Kit, following the manual protocol, automating the extraction using the Biomek i7 Hybrid Workstation, and semi-automating the extraction using the KingFisher Duo Prime Sample Purification System.

This link directs to an external Application Note. This application note compares workflows and yield for the extraction of cfDNA using Apostle MiniMax High Efficiency cfDNA Isolation Kit, following the manual protocol, automating the extraction using the Biomek i7 Hybrid Workstation, and semi-automating the extraction using the KingFisher Duo Prime Sample Purification System.

This link directs to an external poster presented at ASHG 2019. This poster presents the data associated with correlation between mutations found In FFPE tumor tissue and paired cfDNA samples.

This link directs to an external poster presented at ASHG 2019. This poster presents the data associated with the comparison between mutation profiles of paired whole blood and cfDNA samples.

This link directs to an external poster presented at AACR 2019. This poster presents a comparison of matched FFPE and plasma samples to determine how many mutations are seen in both tissues.

This link directs to an external poster presented at AGBT 2019. This poster presents a novel cfDNA extraction kit and show its compatibility with extractions from 200 µl – 5 mL.


Example Data

cfDNA is a group of highly fragmented DNA molecules, with major peak at ~170bp, doublet peak at ~340bp, triplet peak at ~510bp, and so on. Therefore, cfDNA isolation kit capable of highly efficient cfDNA isolation spanning wide cfDNA size distribution is desired. Apostle MiniMax High Efficiency cfDNA Isolation Kit meets such need as demonstrated from its >95% recovery of DNA ladder. This is further validated through isolation of natural cfDNA from human plasma (Exhibit 1A) and urine samples (Exhibit 1B), where Apostle MiniMax High Efficiency cfDNA Isolation Kit offers superior cfDNA isolation efficiency over wide range, specifically covering the 170bp, 340bp, and 510bp cfDNA peaks, when compared with major alternative product.  Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit enables superior and consistent performance of DNA mutation detection, validated by qPCR (Exhibit 2).

Exhibit 1. Superior cfDNA isolation efficiency in human plasma and urine.

A) Cell-free plasma was separated from blood samples by centrifugation for 10 minutes at 2000g at 4oC, then centrifuged for 10 minutes at 16000g at 4oC. cfDNA was isolated from 4mL plasma with Apostle MiniMax High Efficiency cfDNA Isolation Kit (red curve) and major alternative product (blue curve). The isolated cfDNA was characterized by Bioanalyzer 2100. 

B) Cell-free urine was prepared by centrifugation for 10 minutes at 16000g at 4oC. cfDNA was isolated from 20mL urine with Apostle MiniMax High Efficiency cfDNA Isolation Kit (red curve) and major alternative product (blue curve). The isolated cfDNA was characterized by Bioanalyzer 2100. Apostle MiniMaxTM High Efficiency cfDNA Isolation Kit offers superior cfDNA isolation efficiency for both plasma and urine samples.

Exhibit 2. Superior performance of DNA mutation detection isolated with Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit

20 uL of DNA fragment containing the EGFR c.2573T>G L858R mutation (synthetic, ~170 bp), with concentration of 1 ng/uL, 0.1 ng/uL, 0.01 ng/uL, 0.001 ng/uL, was spiked into 1mL TE buffer (blue) or Serum (red) respectively. The mutated DNA fragment was isolated with Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit (Standard Edition), with a final elution volume of 20 uL. qPCR was performed using 1 uL of the isolated DNA, and compared with 1 uL of the corresponding original mutated DNA solution at 1 ng/uL, 0.1 ng/uL, 0.01 ng/uL, 0.001 ng/uL. 

A) Amplification plot showing highly overlapping curves for mutated DNA fragment isolated with Apostle MiniMaxTM High Efficiency Cell-Free DNA Isolation Kit and original DNA solution at different concentrations. 

B) qPCR standard curve generated using original mutated DNA solution, in order to quantify the recovery of DNA isolated with Apostle MiniMax High Efficiency Cell-Free DNA Isolation Kit. DNA isolation recovery rate was calculated to be >90%. 

Note: Displayed DNA concentration series at 1, 0.1, 0.01, 0.001 ng/ul are the concentrations of the original DNA dilution series before spiking into 1mL of serum. The corresponding DNA isolation working concentrations are 20 pg/ul, 2 pg/ul, 0.2 pg/ul, 0.02 pg/ul, respectively.

Applications

Apostle MiniMax technology has been applied in world-class research and development projects of novel liquid biopsy technologies.  Some examples include: 

Multiplex assays, involving the simultaneous use of multiple circulating tumor DNA (ctDNA) markers, can improve the performance of liquid biopsies so that they are highly predictive of cancer recurrence. We have developed a single-tube methylation-specific quantitative PCR assay (mqMSP) that uses 10 different methylation markers and is capable of quantitative analysis of plasma samples with as little as 0.05% tumor DNA. In a cohort of 179 plasma samples from colorectal cancer (CRC) patients, adenoma patients, and healthy controls, the sensitivity and specificity of the mqMSP assay were 84.9% and 83.3%, respectively. In a head-to-head comparative study, the mqMSP assay also performed better for detecting early-stage (stage I and II) and premalignant polyps than a published SEPT9 assay. In an independent longitudinal cohort of 182 plasma samples (preoperative, postoperative, and follow-up) from 82 CRC patients, the mqMSP assay detected ctDNA in 73 (89.0%) of the preoperative plasma samples. Postoperative detection of ctDNA (within 2 wk of surgery) identified 11 of the 20 recurrence patients and was associated with poorer recurrence-free survival (hazard ratio, 4.20; P = 0.0005). With subsequent longitudinal monitoring, 14 patients (70%) had detectable ctDNA before recurrence, with a median lead time of 8.0 mo earlier than seen with radiologic imaging. The mqMSP assay is cost-effective and easily implementable for routine clinical monitoring of CRC recurrence, which can lead to better patient management after surgery. 

We developed a computational program whereby available SD regions can be processed and analyzed efficiently for their potential use as biomarkers of the aneuploidy of interest. For the five common aneuploidies, i.e., trisomy 13, 18, 21, and two sex chromosome aneuploidies, a total of 21,772 candidate SD biomarker sequences together with their corresponding primer/probe sets were generated. The primer/probe sets were tested using a real-time PCR-based multicolour melting curve analysis for simultaneous detection of the five common aneuploidies, and yielded 100% clinical sensitivity and 99.64% specificity when subjected to a clinical evaluation. Following the observations that the SD biomarkers for aneuploidy could be better detected by digital PCR with improved accuracy, we established a noninvasive prenatal testing protocol for trisomy 21 and attained 100% concordance with next generation sequencing.

Our study confirmed that SD regions are preferred biomarkers for aneuploidy detection and in particular SD-based digital PCR could find potential use for NIPT of trisomy. A similar strategy can be applied to other chromosomal abnormality and genetic disorders.

What our community say

(Figure 4) "Most notably, the Apostle particles outperformed all others, achieving almost 2-fold higher recovery yields than the particles supplied in the X kit".
- An independent study by scientists at Johns Hopkins University and University of Pittsburgh (read article)
"In this study, we administered a recombinant bicistronic adeno-associated virus (AAV8) vector coding for both the light and heavy chains of the potent broadly neutralizing HIV-1 antibody VRC07 (AAV8-VRC07) to eight adults living with HIV... Plasma AAV8-VRC07 plasmid DNA was measured by extracting DNA from plasma,... DNA was extracted from serum using an Apostle MiniMax High Efficiency cfDNA Isolation Kit, following the manufacturer’s protocol with slight modification."
- An independent study by NIH, Medical University of South Carolina, MIT and Harvard, Caltech, Duke, Vaxart, and ModeX (read article)
(Figure 1) "For each tube the Apostle MiniMax extracted higher total yield of DNA".
- An independent study by scientists at Beckman Coulter Life Sciences (read data sheet)
"The combination of mutation profiling and CNA profiling can improve the detection of MRD, which may help optimize the treatment strategies for patients with locally advanced rectal cancer (LARC)...cfDNA was extracted from 1.5-4.5 mL of plasma with the Apostle MiniMax cfDNA isolation kit (C40605, Apostle; San Jose, CA, USA). "
- An independent study published in EBioMedicine from Peking Union Medical College, Sun Yat-sen University Cancer Center, Jinchenjunchuang Clinical Laboratory, China
"...This includes recognizing that an innovative product from Apostle is the best technology to address these needs."
- Steve Wowk, Director of Genomics, Beckman Coulter Life Sciences
"The mqMSP assay is cost-effective and easily implementable for routine clinical monitoring of CRC recurrence, which can lead to better patient management after surgery... Plasma DNA extraction was performed using 2 to 5 mL of plasma with the Apostle MiniMax High-Efficiency cfDNA Isolation Kit, according to the product manual. "
- An independent study published in PNAS from Department of Gastroenterology, The First Affiliated Hospital of Wenzhou Medical University, China
"Our study confirmed that SD regions are preferred biomarkers for aneuploidy detection and in particular SD-based digital PCR could find potential use for NIPT of trisomy... The cfDNA was isolated from 1 mL of each plasma sample by the Apostle MiniMax™ High Efficiency cfDNA Isolation Kit (Apostle Inc, San Jose, CA). "
- An independent study published in EBioMedicine from State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Xiamen University, China
"BGI Genomics announces that its Stool Sample Collection Kit, DNA Isolation Kit together with Sample Pretreatment Kit for Methylation Detection have been CE marked. " (Note by Apostle: Apostle is the Original Equipment Manufacturer or OEM for the Stool DNA Isolation Kit mentioned in this news. )
- BGI

Other Information

Apostle also partners with Beckman Coulter to offer the Apostle MiniMaxTM High Efficiency Isolation Kit. If you wish to order the kit through Beckman Coulter, please visit Beckman Coulter's website:

Apostle image 
https://www.beckman.com/reagents/genomic/dna-isolation/from-plasma

For more information about this partnership: https://www.beckman.com/news/liquid-biopsy-partnership-with-apostle

ProductsCat#
Apostle MiniMaxTM High Efficiency Cell-Free DNA Isolation Kit (Standard Edition) (1mL X 10 preps)A17622-10
Apostle MiniMaxTM High Efficiency Cell-Free DNA Isolation Kit (Standard Edition) (1mL X 50 preps)A17622-50
Apostle MiniMaxTM High Efficiency Cell-Free DNA Isolation Kit (Standard Edition) (5mL X 50 preps)A17622-250
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All Publications
A recent independent study conducted by scientists from Johns Hopkins University and University of Pittsburgh, published in a peer-reviewed journal SLAS Technology, demonstrated that, "(Figure 4) Most notably, the Apostle particles outperformed all others, achieving almost 2-fold higher recovery yields than the particles supplied in the X kit". Read the article