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New compositions and methods for cell killing

一种组合物、细胞的技术,应用在植物学设备和方法、用于生物控制的化学品、杀生物剂等方向

Inactive Publication Date: 2014-09-03
OPLON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nothing in any of the aforementioned patents or prior art demonstrates or teaches the cytotoxic effect of solid buffers and ion exchangers through the exchange of protons between the cell membrane and the ion exchanger without adversely affecting the pH of the solution

Method used

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  • New compositions and methods for cell killing
  • New compositions and methods for cell killing
  • New compositions and methods for cell killing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Cytotoxic effect of polyacrylamide gel (PAAG)-coated and uncoated silica beads on Jurkat cells

[0118] Materials and methods

[0119] Uncoated silica beads (~40 nm in size, Sigma, Cat. No. 421553) were in suspension and coated by photopolymerization with polyacrylamide to introduce acidic and basic acrylamide derivatives Silica beads (immobilized electrolyte) were stored in a +4°C freezer until use.

[0120] The acute T cell leukemia Jurkat cell line - clone E6-1 (ATCC number TIB-152) was used. Jurkat cells were maintained in RPMI-1640 medium supplemented with 1 mmol sodium pyruvate, 10% FBS, and penicillin-streptomycin-amphotericin (1:100).

[0121] Viability and microscopy observations

[0122] Add 2 μl of beads (diluted with 0.1% SDS solution) to 10 in 25 μl of PBS 6 in Jurkat cells. LIVE / DEAD dye (LIVE-DEAD viability kit, Molecular Probes) (0.15 μl) was added and incubated at room temperature. Cell morphology and viability were examined using a fluorescen...

Embodiment 2

[0133] Cytotoxic effect of PAAG beads on Jurkat cells

[0134] Materials and methods

[0135] PAAG beads (~500 nm in size) incorporating immobilized electrolyte (immobiline) at various pHs were prepared using standard emulsification techniques. Store the stock solution in a +4°C refrigerator until use.

[0136] The acute T-cell leukemia Jurkat cell line - clone E6-1 (ATCC accession number TIB-152) was used. Jurkat cells were maintained in RPMI-1640 medium supplemented with 1 mmol sodium pyruvate, 10% FBS, and penicillin-streptomycin-amphotericin (1:100).

[0137] Viability and microscopy observations

[0138] Add 2 μl of beads (diluted with 0.1% SDS solution) to 10 in 25 μl of PBS 6in Jurkat cells. Add LIVE / DEAD dye (LIVE-DEAD viability kit, Molecular Probes) (0.15 μl) and incubate at room temperature. Cell morphology and viability were examined using a fluorescence microscope (Axioskop2plus; filters 4-3).

[0139] result

[0140] Microscopy of Jurkat cells treat...

Embodiment 3

[0146] Cytotoxic effect of two Amberlite TM beads CG-120-I and CG-400-II on Jurkat cells

[0147] Materials and methods

[0148] The cytotoxic effect of two Amberlite TM beads CG-120-I and CG-400-II on Jurkat cells was tested. Among them, Amberlite TM CG-120-II (Fluka, 06469) is sulfonic acid functionalized, Na + Form, 200~400 mesh strong acid gel type resin; AmberliteTM CG-400-II (Fluka, 06471) is quaternary ammonium functionalized, Cl - Form, 200-400 mesh strong basic gel-type resin.

[0149] Add 0.15 μl of dye mix (LIVE / DEAD viability kit from Molecular Probes) to 20 μl of Jurkat cells (5 × 10 5 cells). Amberlite TM beads (5 × 10 5 beads) were added to the cell suspension. Immediately transfer 7 μl of the stained cell suspension onto a microscope slide and cover slip. Live and dead Jurkat cells were measured under a fluorescence microscope using a 4-3 green filter.

[0150] result

[0151] The results showed no substantial difference between the control and the...

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Abstract

The present invention discloses an insoluble proton pool or proton source (proton sink or source, PSS), which can be used to kill a living target cell (LTC) by contact or disrupt key intracellular processes of the LTC and / or cell-to-cell interaction. The PSS comprises (i) a proton source or pool providing buffer capacity; and (ii) means providing proton conductivity and / or electrical potential. The PSS effectively disrupts pH homeostasis and / or electrical balance within the defined volume of the LTC and / or disrupts critical cell-cell interactions of the LTC while effectively maintaining the pH of the LTC environment. The present invention also provides a product comprising said PSS and provides an effective method for killing LTC.

Description

technical field [0001] The present invention relates to compositions and methods for killing cells. More specifically, the present invention relates to compositions and methods for killing living target cells or disrupting critical intracellular processes and / or intercellular interactions of said cells while effectively maintaining the pH of said cellular environment. Background technique [0002] Many forms of cells are known to be harmful and potentially fatal to the human body. For example, cancer cells are the second leading cause of death after heart disease in the United States (Boring et al. (1993), CA Cancer Journal for Clinicians 43:7). Cellular microorganisms also cause many diseases. Targeted and selective killing of cells, such as cancer cells and pathogenic bacteria, has been extensively studied in the biotechnology industry. [0003] Microorganisms can invade host tissues and multiply, causing severe disease symptoms. Pathogenic bacteria have been shown to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N61/00A01N25/10A01N25/34A01P1/00A01P15/00A61L2/16A61L27/34A61K45/00
CPCA01N41/04A01N61/00A01N37/20A01N25/34A01N37/08A01N25/10
Inventor 什穆埃尔·比克什潘格莱布·齐尔伯施泰因
Owner OPLON