Aviva Biosciences Corporation

Sakura Finetek U.s.a., Inc.

Douglas T Yamanishi

Lei Wu

Guoliang Tao

Junquan Xu

Mingxian Huang

Jing Cheng

Xiaobo Wang

George T Walker

Jia Xu

Paul G Hujsak

Joe Ouyang

Sara F Snyder

Ping Lin

Charina Schmitigal

Amit D Shah

Andrea Ghetti

Clifford Hom

Wenge Shi

Mengjia Tang

Gioulnar I Harvie

Huimin Tao

David Cerny

Elizabeth Kwok

Charina Yap

Zhaohuai Yang

Jing Tsinghua University Cheng

A method including contacting a tissue or cellular sample with a reagent including an evaporation reducing agent(s) having the general formula X&#x2014;R&#x2014;X, wherein R is an alkyl, alkenyl, alkynyl or an aromatic moiety of 1 or more carbon atoms that may be substituted with an oxygen, nitrogen or sulfur and Xand Xare independently selected to be moiety that is susceptible to hydrogen bonding and processing the tissue or cellular sample. A reagent used in the processing of a tissue or cellular sample with the reagent containing an evaporation reducing agent.

Methods to reduce evaporation during elevated temperature

The present invention recognizes that diagnosis and prognosis of many conditions can depend on the enrichment of rare cells, especially tumor cells, from a complex fluid sample such as a blood sample. In particular, the present invention is directed to methods and compositions for detecting a non-hematopoietic cell, e.g., a non-hematopoietic tumor cell, in a blood sample via, inter alia, removing red blood cells (RBCs) from a blood sample using a non-centrifugation procedure, removing white blood cells (WBCs) from said blood sample to enrich a non-hematopoietic cell, if any, from said blood sample; and assessing the presence, absence and/or amount of said enriched non-hematopoietic cell.

Methods and compositions for detecting non-hematopoietic cells from a blood sample

The present invention includes methods of enriching rare cells, such as cancer cells, from biological samples, such as blood samples. The methods include performing at least one debulking step on a blood sample and selectively removing at least one type undesirable component from the blood sample to obtain a blood sample that is enriched in a rare cell of interest. In some embodiments magnetic beads coupled to specific binding members are used to selectively removed components.

Methods and compositions for separating rare cells from fluid samples

Provided are methods and compositions for isolating and detecting rare cells from a biological sample containing other types of cells, particularly including debulking that uses a microfabricated filter for filtering samples. The enriched rare cells can be used in a downstream process such as identification, characterization or growth in culture, or in other ways. Also included is a method of determining tumor aggressiveness or the number or proportion of cancer cells in the enriched sample by detecting telomerase activity, nucleic acid or expression after enrichment of rare cells. Also provided is an efficient, rapid method to specifically remove red and white blood cells from a biological sample containing at least one of the cell types, leading to enrichment of rare target cells including circulating tumor (CTC), stromal, mesenchymal, endothelial, fetal, stem, or non-hematopoietic cells et cetera from a blood sample.

Methods and compositions for detecting rare cells from a biological sample

The present invention recognizes that diagnosis and prognosis of many conditions can depend on the enrichment of rare cells from a complex fluid sample. In particular, the enrichment of fetal cells from maternal samples, such as maternal blood samples, can greatly aid in the detection of fetal abnormalities or a variety of genetic conditions. In addition, the present invention recognizes that the enrichment of rare malignant cells from patient samples, can aid in diagnosis, prognosis, and development of therapeutic modalities for patients. The invention includes microfabricated filters for filtering fluid samples and methods of enriching rare cells of fluid samples using microfabricated filters of the present invention. The invention also includes solutions for the selective sedimentation of red blood cells (RBCs) from a blood sample and methods of using selective RBC sedimentation solutions for enriching rare cells of a fluid sample. Yet another aspect of the invention is an automated system for processing a fluid sample that includes: at least one filtration chamber that includes a microfabricated filter; automated means for directing fluid flow through at least one filtration chamber of the automated system, and means for collecting enriched rare cells. The present invention also includes methods of using automated systems for separating rare cells from fluid samples. Preferred fluid samples are blood, effusion, or urine samples, and rare cells that can be enriched from such sample include nucleated red blood cells and cancer cells.

Methods, compositions, and automated systems for separating rare cells from fluid samples

The present invention recognizes that diagnosis and prognosis of many conditions can depend on the enrichment of rare cells from a complex fluid sample. In particular, the enrichment of fetal cells from maternal samples, such as maternal blood samples, can greatly aid in the detection of fetal abnormalities or a variety of genetic conditions. In addition, the present invention recognizes that the enrichment of rare malignant cells from patient samples, can aid in diagnosis, prognosis, and development of therapeutic modalities for patients. A first aspect of the present invention is a microfabricated filter for filtering a fluid sample. A microfabricated filter of the present invention comprises at least one tapered pore, and preferably comprises at least two tapered pores whose variation in size is 20% or less. The present invention also includes a method of enriching rare cells of a fluid sample using a microfabricated filter of the present invention. Another aspect of the invention is solutions for the selective sedimentation of red blood cells (RBCs) from a blood sample comprising a red blood cell aggregating agent and at least one specific binding member that selectively binds RBCs. Solutions of the present invention include a combined solution for rare cell enrichment that comprise RBC aggregating agents, at least one specific binding member that selectively binds RBCs, and at least one additional specific binding member for the removal of undesirable sample components other than RBCs. The invention also includes methods of using selective RBC sedimentation solutions and combined solutions for enriching rare cells of a fluid sample. Yet another aspect of the invention is an automated system for processing a fluid sample that includes: at least one filtration chamber that comprises or engages one or more microfabricated filters of the present invention; automated means for directing fluid flow through the one or more filtration chambers of the automated system, and means for collecting enriched rare cells. The present invention also includes methods of using an automated system for separating rare cells from a fluid sample. Preferred fluid samples are effusion, blood, or urine samples, and rare cells that can be enriched from such sample include nucleated red blood cells and cancer cells.

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Carlsbad, CA, United States of America

Douglas T Yamanishi