Show simple item record

dc.contributor.advisorWeitz, David A.
dc.contributor.authorDing, Ruihua
dc.date.accessioned2019-12-17T11:13:47Z
dash.embargo.terms2018-11-01
dc.date.created2017-11
dc.date.issued2017-09-13
dc.date.submitted2017
dc.identifier.citationDing, Ruihua. 2017. A Microfluidic Platform for Rapid Isolation of Cells Based on Secretion of Antigen Specific Antibodies. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences.
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:42061470*
dc.description.abstractDroplet microfluidics provides compartmentalization of cells into picoliter-sized droplets, enabling economical, sensitive and massively parallel biological assays. Droplets can be actively manipulated for different applications. In particular, droplet sorting allows rapid isolation of individual desired droplets from a large excess of undesired droplets. Droplet sorting is particularly powerful when combined with in-droplet assays to identify cells that secrete molecules of interest. However, the accuracy of droplet sorting is limited due to droplet coalescence. In this dissertation I present a droplet filter design that effectively separates droplets by size. The droplet filter has a sharp size cut-off and can distinguish droplets differing in volume by as little as 20%. A simple model explains the behaviour of the droplets as they pass through the filter. We apply the filter upstream of a droplet sorter and show it improves sorting efficiency. We also develop a droplet-based system to accurately identify and isolate individual cells that secrete antigen-specific antibodies. This system eliminates the inefficient and time-consuming steps of cell fusion and clonal expansion required in standard monoclonal antibody generation methods. I present use of this system, coupled with filter-improved droplet sorting, to isolate cells that secrete antigen-specific antibodies. The platform is capable of screening a million droplets within one day and is able to isolate rare antigen-specific cells that consist of 0.1% of the total population. We apply this system to frozen primary rat cells. After sorting, we perform single-cell RT-PCR to identify the IgG-encoding sequences from each isolated cell. A novel ELISA assay is used to verify the target-binding ability of the encoded IgG molecules.
dc.description.sponsorshipChemistry and Chemical Biology
dc.format.mimetypeapplication/pdf
dc.language.isoen
dash.licenseLAA
dc.subjectMicrofluidics
dc.subjectAntibody
dc.titleA Microfluidic Platform for Rapid Isolation of Cells Based on Secretion of Antigen Specific Antibodies
dc.typeThesis or Dissertation
dash.depositing.authorDing, Ruihua
dash.embargo.until2018-11-01
dc.date.available2019-12-17T11:13:47Z
thesis.degree.date2017
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.levelDoctoral
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
thesis.degree.nameDoctor of Philosophy
dc.contributor.committeeMemberXie, Xiaoliang
dc.contributor.committeeMemberCohen, Adam E.
dc.type.materialtext
thesis.degree.departmentChemistry and Chemical Biology
thesis.degree.departmentChemistry and Chemical Biology
dash.identifier.vireo
dc.identifier.orcid0000-0002-7388-1887
dash.author.emaildingrh2014@gmail.com


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record