1. Mawi DNA Technologies LLC
Next Generation BioSampling Devices
Dr. Bassam El-Fahmawi
President & CTO
Collect – Stabilize – Concentrate – Transport – Extract – Store:  All in a Single Tube
Mawi DNA Technologies LLC

2. About Mawi DNA Technologies
Mawi DNA Technologies LLC: Established in Newark, California in 2012
iSWAB: 9 years of dedicated R&D from concept to commercialization
100% US-Made and Manufactured technology.
Maryland
California

3. Mawi DNA Technologies Mission Statement:
Improving human and animal health assessment by enabling efficient and cost effective biological sample collection, room temperature stabilization and transport

4. Genomics Sample Collection Workflow
Transport / Store
Extract / Purify DNA
Analyze

5. Invasive vs. Non-Invasive Collection
Collect cell samples
Stabilize sample in proprietary alcohol-free buffer
Remove and discard swab
Blood
Painful
Requires expertise
Expensive handling infrastructure (cold chain transport and storage)
iSWAB-DNA Collection Device
Painless
Enables self or assisted collection
No cold chain involvement

6. Minimal storage foot print
iSWAB vs. Blood
iSWAB requires less collection time and no professional training ($80-$150)
Vs.
Self collection
Requires specialized personnel
iSWAB does not require cold transport
Ambient shipment in standard mail
Specialized & refrigerated transport
iSWAB can be stored in supplied sample vial >1 year at room temperature
Room Temperature storage vs. Refrigerated Specialized Storage
Minimal storage foot print

7. iSWAB™ Collection Devices Vs
iSWAB™ Collection Devices Vs. Other Non-Invasive Sample Collection Devices

8. Disadvantages of Traditional Non-Invasive Collection Methods
Saliva/Mouthwash
Oral Swabs
Self collection, large volume of saliva required (1-2 mL) can take up to 45 mins
limited to individuals capable of producing sufficient saliva (drugs that cause dry mouth as a side-effect can compromise collection)
Collection solution contains alcohol (requires special extraction requirements and can possibly shear DNA)
Yields sheared, short-fragmented DNA not optimal for complex downstream genomic applications
Up to 40% bacterial content
Readily available in any clinic, simple and easy to use
Low gDNA recovery (up to 2 µg/Sample) with high bacterial contamination
High resampling rate reduces operational efficiency and increases processing costs
Transit time of dry swabs exponentially accelerates sample degradation and reduces DNA recovery substantially
Swab removal and extraction process not automation-friendly

9. iSWAB vs. Saliva Collection
iSWAB wins on Collection Time
Swabs need to be purchased separately with children kits
All collection accessories included
iSWAB-DNA < 5 minutes
Oragene
iSWAB wins on Transportation- Bar-coding and Leak Safe
Secure with O-ring
Transport vessel included
Bar-code comes standard for traceability
No O-ring: Leakage Potential
Transport vessel needed
Bar-coding does not come standard
iSWAB wins on Storage real-estate and cost
1/4th space needed vs. Oragene

10. iSWAB-DNA: Reinventing the swab Patent Pending
Buccal cells collection and concentration device is bar-coded for traceability & chain of custody
Innovative packaging designed to be tamper proof and a sample shipper box

11. Advantages of the iSWAB Non-Invasive Sample Collection Device
Proprietary tube insert design and alcohol-free buffer yields up to 30mg of double-stranded, long fragment DNA ideal for complex downstream genomic applications
iSWAB technology eliminates the need for swab retention
Collects & concentrate predominantly buccal cells thus providing higher quality non-fragmented gDNA
Less than 1% bacterial genomic DNA contamination results in pure sample DNA
Self collection or assisted collection in less than 5 minutes. Suitable for all population segments including infants, toddlers, and elderly
Swab-free sample transport decreases sample processing time without compromising sample integrity
Long-term room temperature stability reduces sample storage and transport costs by eliminating cold chain requirements
Manual and automation friendly tubes for scalability and cost efficiencies

12. Why Buccal Cells not Saliva?
Buccal cells: best surrogate tissue for brain DNA methylation studies
Buccal Cells Are the Dominant Cell Type in The ISWAB
Saliva Content:
Degraded cells
WBCs
High Bacteria Content
Food particles
Mucus
nasal execrations
iSWAB Content:
Buccal Cells
Low Bacteria Content
Cell Types in ISWAB Post Collection:
Total cell counts were assessed by a haemocytometer and differential cell counts by two observers from on Giemsa stained slides. The results are expressed a percentage of each cell type from the total number of cells in the iSWAB collected sample (n=10).

13. iSWAB-DNA Collection Process
Accommodates extraction of cells from multiple swabs per patient
Allows sample concentration
Unique design eliminates the need for swab transport
Self or assisted collection, accommodates all age groups and population segments
Up to 30µg of double stranded, long-fragment human gDNA with <1% bacterial content
2-4 Minutes Collection
iSWAB Sample Collection Process

14. iSWAB-DNA Collection Confirmation
4 Swabs

15. Lab Processing: Saliva vs. iSWAB
Heat at 50C for an hour
Specialized gDNA Extraction Procedures
Compatible with any commercial Blood gDNA extraction kits with no modifications
Faster sample processing ….. Faster TAT…….. $2-4$ saving per sample prep

16. Efficiency of iSWAB Sample Collection & Concentration Vs. Saliva
Comparison of Average Yield of DNA from iSWAB-DNA
&
Oragene Saliva collection methods (OG-500 & OCR-100)
Number of Samples = 48
Average OD 260/280
DNA Concentration (µg/Sample)
iSWAB efficiently suspends and concentrate DNA from buccal cells
iSWAB DNA yield and quality is comparable to Saliva and Superior over standard swab collection
*DNA extraction was performed with DNeasy. DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used in QPCR to relatively quantitate the levels of Human genomic DNA

17. Low Bacterial Genomic DNA Contamination
Comparison of Average Bacterial Genomic DNA Contamination Detected in Different Non-Invasive Sample Collection Methods
Percentage of Bacterial Genomic DNA Content/ Prep
iSWAB bacterial DNA content was determined by RT-PCR using primers specific for the 16S ribosomal RNA gene (n=24)
The bacterial content from iSWAB was <1%, as compared to the other collections methods for the bacterial as reported in literature.
iSWAB Proprietary cell lysis stabilizing solution allows for minimal to negligible bacterial DNA contamination

18. Collection Time and Age Affect on gDNA Yields From iSWAB

19. The Effect of the Sample Collection Time on DNA Yields
 32%
 32%
DNA Concentration (µg/Sample)
DNA extraction was performed with Gentra Puregene Blood extraction kit (n=6):
Each volunteer was asked to self collect Buccal cells using the iSWAB device at different time intervals during the day. Volunteers where asked to abstain from eating, chewing gum and drinking for at least 30 minutes before colleting the samples. Early morning sample was collected first thing in the day and before brushing their teeth. Yields where lower by 32% in midday and 20% in the evening from early morning collections.
*DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used to relatively quantitate the levels of Human genomic DNA .

20. ISWAB is an Efficient Non-Invasive Sample Collection for Various Population Segments
DNA Concentration (µg/Sample)
DNA extraction was performed with Gentra Puregene Blood extraction kit (n=34/age group):
Age groups over 14 years old has performed self collection, where the collection for younger age groups were performed by one of the parents. Sample collection was performed regardless of the time of the day.
*DNA yields where confirmed with 3 different methods: Nanodrop, picogreen assay and primers specific for the human beta actin gene were used to relatively quantitate the levels of Human genomic DNA

21. iSWAB/Saliva/Blood Sample Comparison
Objectives:
Customer was looking for a convenient  DNA collection device that could be easily shipped without refrigeration and be used in clinical DNA sequencing
Conclusions:
iSWAB DNA performed As Well or Better than blood and better than saliva
Sequence analysis of all samples revealed no significant difference between sample types
iSWAB extracted DNA is usable for carrier screening assay
The proportional increase of PCR sensitivity to the gDNA input from iSWAB-DNA device is an indication of very limited PCR inhibitor carryover

22. iSWAB-DNA Performance in NGS: MiSeq Run
FastQ
High Alignment, Minimal bacterial DNA Contamination

23. iSWAB-: Collecting and Preserving DNA Evidence at the Point of Collection
Touch DNA & Reference Samples

24. ISWAB-ID Compatibility with Direct PCR- ID Profiling Kits
ABI Identiflier Direct PCR STR kit

25. ISWAB-Blood for the Blood Drops Collection
Collect – Concentrate – Transport – Store:  All in a Single Tube
Launch Timeline: ASHG – Oct 7th, 2015
MAWI DNA Technologies LLC

26. Current Blood Collection Methods
Venous Blood
Dry Blood Spot
Painful (invasive)
Large volume blood collected
(5-10 mL)
Requires expertise
Expensive handling infrastructure (cold chain transport and storage
High gDNA yields with >23 Kb fragment sizes
Finger brick (minimally invasive)
Five blood spot collected or more
Proper collection requires the blood drop
to go through both sides of the paper
within designed circle
Three hours dry time before transport or
storage
Low yields and highly fragmented DNA

27. ISWAB- Blood Collection Process
Two Sterile Cotton Swabs
Finger disinfectant pad
Lancet to obtain blood from your finger
Carrying case for Lancet and its accessories
ISWAB-Blood Tube
ISWAB-Blood Tube Rack of 50
ISWAB-Blood Collection Instruction Video

28. ISWAB-Blood Vs. Dry Spot Cards

29. High Molecular Weight gDNA From ISWAB-Blood Spots
Agarose gel (0.8%) electrophoresis of gDNA samples isolated
from 100 µL of Human blood collected with ISWAB-Blood Spot using QiaAMP Blood kit. 4 µL of 100 µL elute was used for electrophoresis.
M:λDNA/Hind III+EcoR I

30. Real Time Stability Testing at Room Temperature:kb
23.1
9.4
6.5
4.3
2.3
2.0
0- days
7- days
30- days
45- days
180- days
DNA extraction was performed with QiaAMP Blood extraction kit
Pooled ISWAB-Blood samples (n=24) was subjected to actual non-refrigerated airfreight sample shipping conditions, then stored at room temperature. An aliquate of 200 µL from the pooled sample was processed at each time point with QiaAMP Blood mini (whole blood protocol), then 2 µL of 100 µL elute was used for electrophoresis.
Human HMW gDNA integrity is maintained and nuclease activity neutralized over the real time testing period of 6 months

31. Long Term Stabilization of Blood Analytes

32. ISWAB-Blood Advantage
No dry time required, 5 minutes collection process
After collection, the sample ius ready for room temperature shipment and long term storage
Unlike FTA or FTA like cards, extracting DNA or measuring blood analytes does not require any front prep and/or card punching
DNA extraction can be performed with standard whole blood purification protocol (2- 8 µg of high MW gDNA)
Easy access to the sample for screening and/or in depth analysis purposes (genomics & metabolmics) – reducing resampling and assay failures
Manual and automation compatible with no pre-prep process

33. iSWAB Automation Compatibility

34. ISWAB-DNA Genomics Workflow Compatibility
iSWAB has been successfully tested on various genetic analysis platforms for human, animal, or forensic samples
iSWAB-DNA
No DNA Purification Required
DNA purification recommended -
Extraction can be performed using the “whole blood” protocol of most commercially available kits
qPCR
Sanger Sequencing
NGS
Direct PCR & RT-PCR
Microarrays

35. DNA from iSWAB is Ideal for Simple & Complex Downstream Analysis

36. iSWAB Customizable Solutions
We offer a number of flexible options for customization
Labeling and Packaging
Your branded labels on our box and/or collection devices
Customized return address labels
Include your forms in our package
Custom barcodes
Custom Tube Components
Our patented squeezing insert can be custom molded to fit your vial of choice, and accommodate any size swab head.
We can accommodate different caps including pierceable caps for automation or specific decapper-compatible.
For inquiries please contact us at mawidna.com/distributors

37. ISWAB Product Offering
iSWAB-DNA
iSWAB-RNA
iSWAB-Protein (For drugs screening and protein biomarkers)
iSWAB-Vet / Animals
iSWAB-ID (Foresncis and Paternity testing)
Coming Soon
Oct 7th, iSWAB-Blood (for collecting blood drops from newborn babies)
Q iSWAB-CELLS (Collecting and transporting viable cells up to 4
weeks at RT) – Regenerative medicine, metagenomics, OMICS,
Microbiome , & Cell lines transport
Q MRT-Blood (Whole blood transport with 15 days stability)

38. Reinventing Non-Invasive Sampling!
Thank you!