Kat herine Gat es 5 th year VS TP Leigh Griffit hs Lab American - - PowerPoint PPT Presentation

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Oct 19, 2023 250 likes •412 views

Kat herine Gat es 5 th year VS TP Leigh Griffit hs Lab American Heart Associat ion (AHA) est imat es t hat 2.5% of t he populat ion afflict ed by valve disorders 20,000 deat hs annually 100,000 valve replacement s in 2015 alone

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Kat herine Gat es 5th year VS TP Leigh Griffit hs’ Lab

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 American Heart Associat ion (AHA) est imat es t hat 2.5%

f t he populat ion afflict ed by valve disorders

 20,000 deat hs annually  100,000 valve replacement s in 2015 alone

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 Bovine Pericardium (BP)

 Glutaraldehyde fixation crosslinks and “ hides”

antigens

 10 year lifespan  ~5 years in j uvenile patients  Calcification and destruction  dysfunction

f valve due to residual antigenicity

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 Anything that stimulates an immune response  33 known antigenic proteins in BP  But is it all of them?

Antigen identification is the first step in generating better biomaterials

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 Hypothesis 1: Affinity chromatography can

be used to rapidly identify antigens that human patients have developed as a graft- specific humoral immune response towards glutaraldehyde-fixed BP heart valve replacements.

 Specific aim 1: Apply affinity

chromatography shotgun proteomic antigen identification to determine which BP proteins are xenoantigenic following heart valve replacement.

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 Patient S

election  Protein Extract  Affinity Chromatography  LC-MS / MS  S tatistical Analysis

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Bovine : 39 Mechanical : 44

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 Hypothesis 2: Human graft-specific immune

responses towards BP heart valve implants are independent of carbohydrate epitopes

 Specific aim 2: Apply affinity

chromatography shotgun proteomic antigen identification process to deglycosylated proteins.

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 Patient S

election  Protein Extract 

Deglycosylation/Alpha-galactosidase 

Affinity Chromatography  LC-MS / MS  S tatistical Analysis

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 Extraction solution does not

interfere with deglycosylation process

 Optimal time is 24 hours of

deglycosylation at 37C for non- denatured proteins

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 Valve Center at Mayo Clinic

 Prospective collection

 In Vivo study  Heart Transplant Patients

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BIG PICTURE

By knowing what makes a biomaterial antigenic Y

u can construct something that

isn’t antigenic

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Antigen identification is the first step in generating better biomaterials

BIG PICTURE

By knowing what makes a biomaterial antigenic Y

isn’t antigenic

Questions?