IIT Delhis DIABETES Urge to Cater Insulin HYPERTENSION Renin - - PowerPoint PPT Presentation

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IIT Delhis DIABETES Urge to Cater Insulin HYPERTENSION Renin - - PowerPoint PPT Presentation

Oct 17, 2023 226 likes •525 views

IIT Delhis DIABETES Urge to Cater Insulin HYPERTENSION Renin Inhibitor Vision Develop system utilizing recombinant bacteria to detect the levels of elicitor and producing and releasing required amount of peptidal drug into

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IIT Delhi’s

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Renin Inhibitor Insulin DIABETES HYPERTENSION

Urge to Cater

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Vision

“Develop system utilizing recombinant bacteria to detect the levels of elicitor and producing and releasing required amount of peptidal drug into continuous stream ”

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Key Elements Organism?? Extracellular protein??

Flow system?

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E.coli

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Key Elements E.coli Extracellular protein??

Flow system?

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α-hemolysin system

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Key Elements E.coli α-hemolysin system

Flow system?

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Schematic Vision

Immobilized bacterial system Semi-permeable membrane

Flow System/Vein Bypass

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Key Elements E.coli α-hemolysin system

Continuous Flow Schematic

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B B D D A Peptidal drug Cytoplasm Periplasm

Part 1 – Engineering E. coli to produce and secrete the Peptidal drug

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Part 2 – Designing and optimizing a suitable flow system

1 2 3 Immobilized cells

Peristaltic pump

Feed Tank

Sample collection and testing

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  • 1. Perfusion Reactor

Media in Media + drug out Immobilized cells Perfusion membrane

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Part 2 – Designing and optimizing a suitable flow system

1 2 3 Immobilized cells

Peristaltic pump

Feed Tank

Sample collection and testing

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  • 2. Hollow Fiber Reactor

Hollow fiber with immobilized cells

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Part 2 – Designing and optimizing a suitable flow system

1 2 3 Immobilized cells

Peristaltic pump

Feed Tank

Sample collection and testing

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  • 3. Dialysis Bag

Dialysis bag Holding string Immobilized cells

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LacI IPTG RNA Pol

What exactly does such an idea entail?

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If we waited until we could do something so well that we would not make a mistake, then we will wait forever –

from ‘Organic Chemistry’ by IL Finar

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hlyB and hlyD:- The Trans membrane components

The hemolysin CABD operon

hlyA:- The evil toxin hlyC:- The instigator

B B D D

Cytoplasmic side Periplasm

hlyA tag

yfp

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α-hemolysin system

After Ivaylo Gentschev,Guido Dietrich and Werner Goebel (2002). Trends Microbiol. 10, 39-45.

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  • Chimeric PCR used to create

protein fusion.

  • PCR products digested

and ligated in two steps with pLac and various RBS sequences.

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  • Reporter construct for study of secretion

characteristics submitted(BBa_K408000).

  • Biobricks necessary for extracellular secretion
  • f proteins created( not yet submitted).
  • The flow system analysis is ready to proceed
  • nce the cellular machinery is assembled.
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Future Directions

  • Need to improve the efficiency of transcriptional

control

  • Study the possibility of controlling the expression
  • f the trans membrane components for faster

response time.

  • Hope to combine this with a sensor apparatus to

create a targeted device

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Acknowledgments

Our Mentors

  • Dr. D.Sundar
  • Dr. Atul Narang

Our Graduate Advisors

  • Ms. Somya Mani
  • Mr. Abhinav Grover

Special Thanks to Prof. Agneta-Richter Dahlfors and

  • Dr. Peter Kjall from the Karolinska Institute, Sweden.

Our Sponsors Indian Institute of Technology, Delhi

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Thank you…..