. .

Hydrogel bead assay for screening in Droplet Microfluidics

Hydrogel bead assay for screening in Droplet Microfluidics

By In Aiv Internship On November 25, 2019

Internship title: Hydrogel bead assay for screening in Droplet Microfluidics

Name: Unit Antibody in Therapy and Pathology
Affiliation: Institut Pasteur – INSERM U1222
Address: 25 rue du Dr Roux
E-mail: angga.perima@pasteur.fr

LAB Director
Name: Pierre BRUHNS
Phone number: +33(0)145688629
E-mail: pierre.bruhns@pasteur.fr

Name: Angga Perima
Phone number: +33(0)144389183
E-mail: angga.perima@pasteur.fr

Subject Keywords: 1. droplet microfluidics
2. hydrogel beads
3. polymerization
4. antibody
5. antigen
Tools and methodologies: 1. beads synthesis
2. laser detection
3. ELISA sandwich
4. RNAseq
Summary of lab\’s interests:
Project summary: Currently, we use assays that can detect cells secreting specific IgG to a determined antigen/target. These Antibody Secreting Cells (ASC) are isolated from mice immunized against that antigen and screened using droplet based microfluidics technologies1. The current assay is based on magnetic beads forced to align into a “beadline” to create a physical surface inside the droplet. This surface serves as the basis of a fluorescent sandwich ELISA: the beads capture secreted antibodies, and only antigen-specific antibodies capture fluorescent antigen that is present in soluble form in the droplet. Based on the fluorescent signal detected for the beadline, droplets containing ASC of interest can be sorted and proceed for RNA sequencing2 by various methods. Our aim is to obtain the sequences corresponding to the genes encoding antibodies to establish antibody repertoires specific for an antigen.

We aim to develop novel assays without magnetic beads that can be applied for droplets screening in microfluidic chips as well as in flow cytometry instruments (FACS). The aim is to sort cells secreting an antibody specific to certain antigens. We wish to develop assays using hydrogel beads as a physical surface to trap antibodies and antigens. This assay should be compatible with subsequent RNA barcoding inside the droplet, followed by bulk RNA sequencing.

This project is multidisciplinary (biology, physics, chemistry, and engineering), and we are looking for a motivated student with chemistry or biochemistry background to work on this project.
Interdisciplinary aspect of the project: 1. bead synthesis (chemistry)
2. droplet microfluidics (physics, chemistry, engineering)
3. sorting specific Antibody Secreting Cells (immunology).
4. RNAseq (molecular biology, bioinformatics)