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“Enabling Be,er Outcomes through Be,er Diagnos7cs” Magne&c nanosensors for sensi&ve protein detec&on Adam Seger, Ph.D. Electrical Engineer

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Page 1: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

“Enabling  Be,er  Outcomes  through    Be,er  Diagnos7cs”  

 

Magne&c  nano-­‐sensors  for  sensi&ve  protein  detec&on  

Adam  Seger,  Ph.D.  Electrical  Engineer  

Page 2: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Nanoengineered  Biomedical  Devices  

•  What  does  it  mean?  •  What  do  we  need?  – A  biologically  relevant  problem  – A  technology  that  can  address  that  problem  

•  How  do  we  get  there?  – Get  engineers  together  with  scien&sts  

Page 3: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

ELISA  

•  Enzyme  Linked  Immunosorbent  Assay  •  Method  established  1971  – Replaced  radioimmunoassay  

•  Chromogenic,  fluorogenic  •  Detect  proteins,  an&bodies,  pep&des…  

Page 4: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

ELISA  

hRp://www.epitomics.com/images/products/sandwich.jpg  

Page 5: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Giant  Magnetoresistance  (GMR)  

•  Discovered  in  1988  – Albert  Fert  and  Peter  Gruenberg  

•  Nobel  Prize  in  2007  

Page 6: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Giant  Magnetoresistance  

hRp://www.nims.go.jp/apfim/GMR.html  

Page 7: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

What  Happens  When  We  Combine  These  Technologies?  

??  

Page 8: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

We  Get  MagArray!  

Simple,  ultra-­‐sensi1ve,  mul1plex  immunoassay  system    

10  mm  12  mm  

Page 9: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

MagArray  System  What  do  we  mean  by:  

•  Simple    -­‐    Small,  portable  system  about  the  size  of  a  &ssue  box…No  moving  parts…No  microfluidics…  Basic,  easy  step  process.      •  Ultra-­‐sensi1ve    -­‐    Have  been  able  to  dis&nguish  concentra&ons  as  low  as  50  aRomolar.    Depending  on  the  target  protein,  typical  detec&on  limits  are  in  the  femtomolar  /  picomolar  range.  • Mul1plex    -­‐    80  individual  sensors  per  chip.    One  chip  per  sample.    Factoring  in  controls  and  replicates,  researchers  typically  analyze  10-­‐20  proteins  per  chip.    

Detection of Multiple Analytes, Each at 1 pg/mL in PBSS. Osterfeld and H. Yu, October-2-2008

000E+0

50E-6

100E-6

150E-6

200E-6

250E-6

300E-6

350E-6

400E-6

0 5 10 15 20 25 30 35 40Minutes

Sign

al, V

p

100:1

75:1

Biotinylated Sensors

1 pg/mL Analytes

Photo of Individually Functionalized Sensors:

Biotin / BSA

GC

SF

Biotin

Epoxy

TNF-α

IL-1α

Eotaxin

INF-γ

LFVEGF

CEA

Page 10: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

MagArray  History  •  Founded  in  2005  by  team  of  Stanford  professors  who  adapted  a  magne&c  nanopar&cle  technology  from  the  computer  disk  drive  industry  for  use  in  medical  diagnos&cs  •  Focus  on  market  need  to  beRer  detect  proteins  and  develop  biomarker  panels    •  Basic  technology  development  from  2007-­‐2011  funded  primarily  by  federal  grants  and  contracts  •  Partnership  with  Aisin  Seiki  in  2011  •  Currently  located  in  life  sciences  incubator  in  Sunnyvale    • Working  with  alpha  customers  on  applica&on  development  

Page 11: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

How  the  MagArray  System  Works  

Sample  added  to  chip  with  pre-­‐func&onalized    capture  an&bodies  

Target  an&gen  binds  to  capture  an&body  

Detec&on  an&body  binds  to    target  an&gen  

Magne&c  nanopar&cle  binds  to  detec&on  an&body  

Magne&c  nanopar&cle  generates  magne&c  field  that  induces  electric  current  in  GMR  sensor  

Page 12: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Real  Time,  Mul&plexed  Results  

Page 13: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Superior  Sensi&vity  and  Dynamic  Range  

Compared  to  ELISA,  the  MagArray  system  was  able  to  detect  CEA  at  a  much  lower  concentra1on  and  across  a  wider  dynamic  range  

Page 14: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Flexible  Assay  Protocols  

【Express  Assay】  •  <  10  minutes  •  Homogeneous  Assay  •  Can  be  run  with  whole  blood  

【Standard  Assay】  •  About  1  hour  •  Excellent  sensi&vity  

Add  Sample  Start  

Add  Detec&on  An&bodies  

Incubate  30min  

Add  Magne&c  Par&cles  

Incubate  30min   Results  2-­‐5  

min  

Add  Sample,  An&bodies,  Par&cles  

Start   Results  2-­‐5  min  

【Enhanced  Sensi1vity】  •  Best  sensi&vity  •  Same  basic  process  •  Ideal  for  low  abundance  proteins  

Add  Sample  Start   Add  Detec&on  

An&bodies  Incubate  4  hours  

Add  Magne&c  Par&cles  

Incubate  4  hours   Results  2-­‐5  

min  

Page 15: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Standard  vs  Express  Assay  Performance  

Page 16: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Minimal  Sample  Volume  

5  µl  samples  

Diluted  1:100  

Ran  EGFR/HE4  in  Duplicate  

Ran  Autoan&bodies  in  Duplicate  

Extra  Vial  for  Repeats  

Page 17: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Wash-­‐Free  Assay  for  Analyzing  Cross  Reac&vity  

Add  magne&c  nanopar&cles,  then  detec&on  an&bodies  sequen&ally:  an&-­‐EGFR,  an&-­‐CEA,  and  an&-­‐EpCAM    

Page 18: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Sequen&al  Addi&on  of  An&bodies  

An1bodies  can  be  added  in  a  stepwise  process  to  iden1fy  and  isolate  cross  reac1vity  issues.    

Page 19: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Advantages  In  addi7on  to  superior  performance  in  sensi7vity,  dynamic  range,  mul7plexibility,  size,  cost,  etc.,  the  following  unique  characteris7cs  are  worth  highligh7ng:  •  Real  Time  Read  Out  –  Sensors  are  scanned  every  4  seconds,  and  results  are  displayed  as  they  

are  generated,  thus  allowing  researchers  to  monitor  binding  as  it  takes  place.    Extremely  useful  for  “go/no  go”  decisions  in  a  fast  paced  environment.  

•  Whole  Blood  –  Op&cal  interference  caused  by  red  blood  cells  is  not  a  factor  for  MagArray’s  magne&c  technology.    While  sensi&vity  is  slightly  less  than  serum,  whole  blood  can  be  used  as  a  suitable  sample  type.  

•  Sequen1al  Addi1on  of  An1bodies  –  MagArray’s  open  well  system  allows  for  detec&on  an&bodies  to  be  sequen&ally  added  at  any  stage  during  the  process.    Researchers  can  iden&fy  and  isolate  cross  reac&vity  issues  in  mul&plex  assays,  as  well  as  design  assay  procedures  to  circumvent  certain  cross  reac&vity  problems.  

•  No  Wash    -­‐    MagArray  sensors  only  detect  magne&c  par&cles  that  have  bound  to  the  analytes  of  interest.    There  is  no  need  to  wash  aler  the  addi&on  of  label.    This  is  of  par&cular  advantage  when  analyzing  low  affinity  proteins  that  tend  to  get  washed  away  during  typical  assay  procedures.    

•  Small  Sample  Volume    -­‐    Assays  can  be  run  using  as  liRle  as  10-­‐20ul.    MagArray’s  system  allows  researchers  to  run  mul&plex  assays  on  minute  samples  from  small  animals  such  as  mice  or  rats.    It  is  also  extremely  valuable  for  limited  human  samples  that  cannot  be  readily  re-­‐obtained.  

 

Page 20: MagArray IEEE Presentation 2013-11-19sites.ieee.org/.../2013/09/MagArray-IEEE-Presentation-2013-11-19.pdf · – Replaced(radioimmunoassay(• Chromogenic,fluorogenic • Detectproteins,

Notable  Papers