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An antibody bound synthetic vesicle containing active agent molecules

An active agent, vesicle technology, applied in the field of antibody-bound synthetic vesicles

Inactive Publication Date: 2011-05-18
BANYAN BIOMARKERS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, TBI has specific issues that previous studies have not properly addressed

Method used

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  • An antibody bound synthetic vesicle containing active agent molecules
  • An antibody bound synthetic vesicle containing active agent molecules
  • An antibody bound synthetic vesicle containing active agent molecules

Examples

Experimental program
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Effect test

example 1

[0064] Example 1: Reagents and Antibody Target Identification: Exemplary materials and reagents used are optionally as follows. It will be appreciated that those of ordinary skill in the art will recognize other reagents that are equally operable to make and use the present invention. Places for obtaining these reagents are also known to those skilled in the art and include, for example, Invitrogen Corp. (Carlsbad, CA), EMD Chemical, Inc., VWR Scientific (West Chester, PA), Santa Cruz Biotechnology (Santa Cruz, CA) and other biological reagent suppliers. Materials and reagents illustratively include: PBS solution, 3.7% formalin solution, Tris-glycine electrophoresis buffer (Invitrogen), gel transfer buffer (Invitrogen), precast gel electrophoresis (Invitrogen), western blot filter paper and Polyvinylidene fluoride (PVDF) membrane (Invitrogen), methanol, Tris-buffered saline (TBST) solution containing Tween 20, (Sigma), rat primary cerebellar granule neurons, skim milk powder,...

example 2

[0067] Example 2: Recognition of NMDA / glycine receptors on the surface of CGN cells by immunocytochemical antibodies

[0068] Antibodies targeting surface expressed NMDA receptors or glycine receptors were confirmed by immunocytochemistry using CGN cells. CGN were grown on coverslips for 7 days. It was then washed with phosphate buffered saline (PBS), fixed with 4% paraformaldehyde solution at 4°C for 10 minutes, and then blocked with 5% normal goat serum in TBST for 30 minutes at room temperature to prevent any non-specific binding of antibodies.

[0069] Divide the cells into two experimental groups. One set was washed with cold methanol for about a minute to open the cell membranes and make the cells permeable to the antibody. In another group, the cell membrane was left intact so that the antibodies could not cross the membrane. Both groups were incubated overnight with primary anti-NMDA and glycine receptor antibody (1 / 500), then washed, and incubated with FITC-seconda...

example 3

[0071] Example 3: Conjugation of Biotin to Antibody

[0072] Antibodies of the invention are biotinylated by methods known in the art. Briefly, antibodies were transferred to 1X modification buffer (100 mM phosphoric acid, 150 mM sodium chloride, pH 7.2-7.4). A biotin solution was prepared at a concentration of 0.5 mg of biotin per 25 µL of DMF (dimethylformamide). Add 0.8 µL of biotin solution to the antibody solution and incubate for two hours at room temperature on a rotary agitator. After incubation, the solution was transferred to a spin-on filter and centrifuged 4 times at 12000 xg for 30 minutes to filter out unbound biotin molecules. The remaining solution that was not filtered was stored at 4°C until future use.

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Abstract

A process is provided of delivering at least one active agent cargo molecule into an neuronal cell whereby a cargo molecule is placed within a synthetic vesicle such as a liposome and a biotinylated protein such as an antibody is bound to the synthetic vesicle to form a protein bound synthetic vesicle whereby the protein recognizes a receptor expressed on the surface of a neuronal cell, and exposing the protein bound synthetic vesicle to the cell until the cargo molecule is delivered into the neuronal cell. Numerous cargo molecules are delivered by the inventive synthetic vesicle including a calpain inhibitor and a caspase inhibitor. The protein illustratively targets a cellular receptor for a ligand such as glutamate, glycine, dopamine, nicotine, muscarine, acetylcholine, or serotonin, and the like.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 61 / 045,748, filed April 17, 2008; which is incorporated herein. technical field [0003] The present invention generally relates to a synthetic vesicle targeted to cells of the central nervous system (CNS) or peripheral nervous system (PNS), in particular an antibody-conjugated synthetic vesicle for delivery of molecules contained within the vesicle to A cell expressing the receptor target of the antibody. Background technique [0004] Approximately 270,000 people experience moderate to severe traumatic brain injury (TBI) each year. TBI is "an acquired injury to the brain caused by an external physical force...resulting in impairment of one or more areas such as cognition; language; memory; attention; reasoning; abstract thinking; judgment; problem solving; sensation, perception and motor ability; psychosocial behavior; physiological function; informa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K9/127A61K47/34A61P25/28
CPCA61K49/0054A61K49/0043A61K49/0058A61K49/0084A61K9/127A61K47/48823C07K16/286A61K47/6913A61P25/00A61P25/28A61P35/00A61P43/00
Inventor 乔纳森·王贾里德·V·古德曼斯蒂芬·弗兰克·拉尔内尔凯文·卡王·王
Owner BANYAN BIOMARKERS INC
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