Unlock instant, AI-driven research and patent intelligence for your innovation.

Methods and systems for screening using microcapillary arrays

A microcapillary, array technology, applied in laboratory containers, chemical instruments and methods, support of flat carriers, etc., can solve the problem of not allowing inactivation and recovery

Pending Publication Date: 2022-01-18
XCELLA BIOSCIENCES INC
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although such techniques provide analytical information about a specific sample (such as the presence and likely amount of a specific biomolecule in solution, or the sequence of a specific nucleic acid or polypeptide), they generally do not allow for the analysis without inactivating the sample of interest or using Recovery of biological samples identified by the assay without otherwise compromising the sample of interest

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods and systems for screening using microcapillary arrays
  • Methods and systems for screening using microcapillary arrays
  • Methods and systems for screening using microcapillary arrays

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0273] Example 1. Screening for secreted EGFR binding proteins

[0274] Figure 1A-Figure 1C An exemplary screening method for soluble proteins capable of associating with a cell surface protein (e.g., epidermal growth factor receptor ("EGFR")) as an immobilized target molecule (in this case, an immobilized target protein) is shown . Figure 1A (Left panel) shows target cells, expressing EGFR on their surface. Also shown is a "library expressing cell" expressing a population of variant proteins and a number of "fluorescent detection antibodies" in microcapillary solution. A bottom view of the microcapillary array is shown in the right panel.

[0275] The composition of each microcapillary determined according to this screen:

[0276] 1. Cells that secrete the variant protein of interest ("library expressing cells"). The target variant protein is preferably a member of a variant protein group, ie a protein library.

[0277] 2. Target protein immobilized on the surface of...

Embodiment 2

[0290] Example 2. Hybridoma Screening for Mammalian Cells

[0291] general background

[0292] Current methods of screening for binding interactions between proteins or other target molecules typically rely on the use of "display" methods such as phage display, bacterial display, yeast display, mammalian display or viral display. In display methods, a library of genes encoding protein variants is expressed on the surface of a cell or phage. Protein variants are incubated with soluble forms of the target molecule to identify protein variants capable of binding the target. Libraries can be screened by panning or by fluorescence activated cell sorting ("FACS"). Such assays have two major limitations: 1) the engineered protein is usually tethered to a display platform; and 2) it is often advantageous to have a soluble form of the target molecule. Therefore, it has been difficult to develop reliable assays for variant proteins binding to many target molecules, especially mem...

Embodiment 3

[0309] Example 3. Yeast library screening for mammalian cells

[0310] To identify the best secreting yeast plasmid vectors, a library of yeast vectors expressing scaffold proteins designed to bind to EGFR on the surface of cancer cells was established. This library contains yeast cells with various soluble expression levels of scaffold proteins. Using the assay, variant expression libraries are screened to recover plasmid vectors that highly express the desired scaffold protein. In this experiment, the secreted scaffold has a c-Myc tag, which can be labeled with a fluorescently labeled antibody.

[0311] Material:

[0312] cell:

[0313] Scaffold Protein Yeast Secretion Library

[0314] A431 cells (a human cancer cell line expressing high levels of EGFR)

[0315] Detection antibody:

[0316] chicken anti-c-Myc

[0317] Anti-chicken secondary antibody labeled with Alexa488

[0318] Medium for cell culture:

[0319] DMEM-10%FBS

[0320] SD-CAA Basic Yeast Medium ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
heightaaaaaaaaaa
diameteraaaaaaaaaa
pore sizeaaaaaaaaaa
Login to View More

Abstract

High-throughput methods for screening large populations of variant proteins are provided. The methods utilize large-scale arrays of micro capillaries, where each micro capillary comprises a solution containing a variant protein, an immobilized target molecule, and a reporter element. Immobilized target molecules may include any molecule of interest, including proteins, nucleic acids, carbohydrates, and other bio-molecules. The association of a variant protein with a molecular target is assessed by measuring a signal from the reporter element. The contents of micro capillaries identified in the assays as containing variant proteins of interest can be isolated, and cells expressing the variant proteins of interest can be characterized. Also provided are systems for performing the disclosed screening methods.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 62 / 830,978, filed April 8, 2019, which is expressly incorporated by reference in its entirety for all purposes. technical field [0003] The present disclosure provides methods and systems for screening using microcapillary arrays. [0004] Background of the invention [0005] Analysis of biological samples, including the identification, characterization, and reengineering of proteins, nucleic acids, carbohydrates, and other important biomolecules, has benefited greatly from the scaling up of sample numbers and the shrinking of sample sizes. For example, two-dimensional microarrays of biological materials, such as DNA microarrays, have enabled the development of high-throughput screening methods involving multiplex methods for processing samples and detecting results. [0006] In some cases, the methods described above benefit from their combination ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00C12N15/10G01N33/68
CPCC12N15/1086C40B40/08B01L3/50857B01L9/52B01L2200/0668B01L2200/16G01N33/5023G01N2333/70596G01N33/6845G01N33/54366G01N33/68G01N33/6803B01L2300/0838B01L2400/0406
Inventor 陈博瑞安·路易斯·凯利谢子咏
Owner XCELLA BIOSCIENCES INC