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My#Role#on#the#Project#

/  Tes2ng#an2bodies#/  Determining#sensi2vity#of#IRIS#for#pseudotyped#VSV#detec2on#/  Determining#repeatability#/  Valida2ng#new#IRIS#instruments#(IRIS#on#med#campus)#/  Standardizing#the#chip#prepara2on/incuba2on#protocol##/  To#improve#sensi2vity#and#repeatability#with#surface#op2miza2on#

Hi, my name is Collaborator X and I'm responsible for optimizing the protocol that we use to coat the chips with antibodies. My job is to make sure that each chip has as few foreign particles as possible, and that each antibody has a high affinity for the virus we are trying to detect. After incubating the chips, I scan them and analyze the data. Then, I try to come up with ways to lower the variability and improve the sensitivity of our results.

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Chip%Prepara*on%Protocol%

Chip%Spo(ng%&%Washing:%%0%An*body%solu*ons%are%spo7ed%at%305%mg/ml%in%PBS%with%50%mM%Trehalose.%%%%%0%ADer%overnight%incuba*on%in%the%spo7er%(at%60%%humidity),%chips%are%washed%in%PBST%for%30%minutes,%rinsed%in%1X%PBS%for%3%minutes%and%dI%for%3%minutes%and%dried%with%argon.%

Incuba4on%of%Chips%with%Concentrated%Samples:%!0%Chips%are%scanned%on%IRIS%for%pre0incuba*on%par*cle%counts.%%%0%Chips%are%incubated%with%1%ml%of%the%virus%sample%for%2%hours%on%shaker.%%

0%ADer%incuba*on%ends,%chips%are%washed%in%1X%PBS%for%3%*mes,%each%1%min%and%then%0.1X%PBS%for%1%min,%dipped%in%dI%for%10%seconds%and%pulled%out%of%the%water%slowly%to%let%the%water%slide%down%the%chip.%%%0%Chips%are%scanned%with%IRIS%for%analysis.%

Spotting and Washing Pre-incubation scan

Incubation with virus Post-incubation scan

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LED

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Future&Work&

*  Dilu.on&experiment&for&Marburg*pseudotyped&VSV&

*  10^4&PFU/ml&detec.on&in&30&minutes&

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*  Tes.ng&of&New&Lassa&An.bodies&(7&of&them)&&&

*  Dilu.on&curve&for&Lassa*pseudotyped&VSV&for&LOD&determina.on&

Obtain dilution curves for!Marburg- and Lassa-pseudotype VSV

Obtain dilution curves for!Marburg- and Lassa-pseudotype VSV

Try to detect viruses at lower !concentrations (104 PFU/ml) after !shorter incubation periods (30 minutes)

Obtain dilution curves for!Marburg- and Lassa-pseudotype VSV

Try to detect viruses at lower !concentrations (104 PFU/ml) after !shorter incubation periods (30 minutes)

Test 7 new Lassa antibodies

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