Revision as of 23:20, 15 October 2018

Design

We used a Phage Display screening to create a biosensor that aggregated around the target molecule with the hypothesis that this would create a larger response.

Phage display is a method that is usually used to find a protein that binds to another protein. However using this technique on a whole eukaryotic organism is very uncommon, and therefore this project represents breaking new ground, albeit with the inevitable cost of hammering out the problems that come with such an undertaking.

In order to make our method work, we had to immobilize our worms and be able to extract the bound phages without also extracting the worms! Since our worms are far larger than a phage, we used an eppendorf filter insert that had pores too small for strongyles to pass through and allowed our phages to bind to these worms.

Using this process, we found several peptide sequences with potential high affinity to our worms. These sequences, after further validation, can in future be used to synthesize E. coli that will bind to our worms!

To read more about the general design of a Phage Display Screening and why we chose to apply this method, click here:

Experiment

There are no readily available protocols for whole-organism exterior screenings using phage display, to read more closely what how we adapted this procedure to fit our needs read more here.

Protocol

For this experiment we used a modified version of the protocol supplied with the kit from New England Biolabs. You can find the original protocol here.

Read more about our modified protocol here.

Results

Our three parallell pannings and subsequent analysis led to the following peptides being characterized:

Table 1: Final peptides and their characteristics. *TUP = Target-Unrelated Peptide.

	Sample	Aligned Sequence	Propagation Advantage	Predicted Polysterene Binder	Known TUP*-Motif
Series 1:	EF01122224: TPIFLPTPAQEH	TPIFLPTPAQEH---	Yes	No	No
Series 3:	EF01122218: FSPTQANTIHRW	---FSPTQANTIHRW	No	No	No
	EF01122220: VGGTVQSESHRR	--VGGTVQSESHRR-	No	No	No
	EF01122222: SMGRTDYVQQLR	-SMGRTDYVQQLR--	No	No	No
	EF01122217: RVQPAHFNVMGQ	--RVQPAHFNVMGQ	No	Yes	No

Notebook

Here you can find our lab notes, the notes and scribbles we took during the course of our experiments:

Notebook Protocol

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