Capture, purification, and release of biological substances using a surface coating

• US 9,541,480 B2
• Filed: 06/28/2012
• Issued: 01/10/2017
• Est. Priority Date: 06/29/2011
• Status: Active Grant

• Current Assignee: Academia Sinica (Government of The Republic of China)
• Original Assignee: Academia Sinica (Government of The Republic of China)
• Inventors: Wu, Jen-Chia, Chang, Ying-Chih, Wu, Han-Chung, Tseng, Po-Yuan
• Primary Examiner(s): Claytor, Renee
• Assistant Examiner(s): Papciak, Sharon M
• Application Number:
  US14/128,354
• Publication Number:
  US 20140255976A1
• Time in Patent Office: 1,657 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 17/14 Peptides being immobilised on, or in, an inorganic carrier
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

This invention relates to a surface coating for capture circulating rare cells, comprising a nonfouling composition to prevent the binding of non-specific cells and adsorption of serum components; a bioactive composition for binding the biological substance, such as circulating tumor cells; with or without a linker composition that binds the nonfouling and bioactive compositions. The invention also provide a surface coating for capture and purification of a biological substance, comprising a releasable composition to release the non-specific cells and other serum components; a bioactive composition for binding the biological substance, such as circulating tumor cells; with or without a linker composition that binds the releasable and bioactive compositions. The present invention also discloses a novel microfluidic chip, with specific patterned microstructures to create a flow disturbance and increase the capture rate of the biological substance.

Claims

1. A method comprising:

(a) contacting a biological sample comprising rare cells and non-specific cells to a non-fouling lipid layer of a microfluidic device, the microfluidic device comprising an antibody which selectively binds to rare cells, and wherein the non-fouling lipid layer is non-covalently associated with the antibody;

(b) binding the rare cells to the antibody;

(c) washing the non-fouling lipid layer with a liquid, wherein the non-specific cells in contact with the non-fouling lipid layer are released from the non-fouling lipid layer; and

(d) flowing a liquid comprising bubbles through a channel of the microfluidic device of step (c), thereby releasing the rare cells from the microfluidic device.

2. The method of claim 1, wherein the released rare cells are viable.

3. The method of claim 2, further comprising collecting greater than 70% of the bound rare cells.

4. The method of claim 3, wherein the collected rare cells remain viable.

5. The method of claim 1, wherein the non-fouling lipid layer comprises a lipid monolayer.

6. The method of claim 1, wherein the non-fouling lipid layer comprises a lipid bilayer.

7. The method of claim 6, wherein air inside the bubbles has high affinity with hydrophobic tails of the lipid bilayer.

8. The method of claim 1, wherein the non-fouling lipid layer comprises liposomes.

9. The method of claim 1, wherein the non-fouling lipid layer is coupled to a solid substrate by a surface linker.

10. The method of claim 1, wherein the non-fouling lipid layer is from 2 nm to 300 μ

m thick.

11. The method of claim 1, wherein the rare cells are circulating tumor cells (CTCs).

12. The method of claim 1, wherein the antibody comprises a heavy chain and a light chain that binds EpCAM, wherein

(a) the heavy chain comprises CDR1, CDR2, and CDR3 of SEQ ID NO:

1,

(b) and the light chain comprises CDR1, CDR2, and CDR3 of SEQ ID NO;

2.

13. The method of claim 1, wherein the non-specific cells are selectively removed from the non-fouling lipid layer through application of the liquid at shear stress of from about 2.5 dyne/cm² to about 10 dyne/cm².

14. The method of claim 1, wherein the liquid comprising bubbles disrupts the non-fouling lipid layer and lifts off a top layer of the non-fouling lipid layer, together with the rare cells and bound antibody.