Papers

A Novel Platform for Detection of CK+ and CK- CTCs

 Conclusion:
 1.   Current assays for CTC capture likely miss populations of cells that have undergone EMT. Capture and study of CTCs that have undergone EMT would allow a better understanding of the mechanisms driving metastasis.
2. The results confirm the utility of our microfluidics platform as a reliable method for assay development and efficient recovery of CTCs. We observed that a potentially important population of cancer cells is present in circulation that would likely be missed by standard detection criteria. 

Chad V. Pecot, Farideh Z. Bischoff, Julie Ann Mayer, Karina L. Wong, Tam Pham, Justin Bottsford-Miller, Rebecca L. Stone, Yvonne G. Lin, Padmavathi Jaladurgam, Ju Won Roh, Blake W. Goodman, William M. Merritt, Tony J. Pircher, Stephen D. Mikolajczyk, Alpa M. Nick, Joseph Celestino, Cathy Eng, Lee M. Ellis, Michael T. Deavers, and Anil K. Sood

Cancer Discovery December 2011 1:580-586; Published OnlineFirst November 3, 2011; doi:10.1158/2159-8290.CD-11-0215  

http://cancerdiscovery.aacrjournals.org/content/1/7/580.full.pdf+html  

Efficient capture of circulating tumor cells with a novel immunocytochemical microfluidic device 

 Conclusion:
 1.   Overall, data concluded that the CEE system conforms to simple physical principles
and that it robustly and efficiently captures tumor cells under a variety of operating conditions.
2.  There is a clear two step recommendation. First, clinicians should employ antibody cocktails with a common capture principle, here biotin, in order to maximize relevant cell capture.  Second, while the data presented here show that in most cases with sufficient surface antibody density, greater than 70% capture was achieved, a channel design which forces variations in flow velocities along the channel length may overcome potential inefficiencies in capture seen at low antigen densities. 

There is a clear two step recommendation. First, clinicians should employ antibody cocktails

with a common capture principle, here biotin, in order to maximize relevant cell capture.18

Second, while the data presented here show that in most cases with sufficient surface antibody

density, greater than 70% capture was achieved, a channel design which forces variations in

flow velocities along the channel length may overcome potential inefficiencies in capture seen

at low antigen densities. 

There is a clear two step recommendation. First, clinicians should employ antibody cocktails

with a common capture principle, here biotin, in order to maximize relevant cell capture.18

Second, while the data presented here show that in most cases with sufficient surface antibody

density, greater than 70% capture was achieved, a channel design which forces variations in

flow velocities along the channel length may overcome potential inefficiencies in capture seen

at low antigen densities. 

Mary Nora Dickson,1,2,a) Pavel Tsinberg,1 Zhongliang Tang,1, Farideh Z. Bischoff,1 Timothy Wilson,1 and Edward F. Leonard2
1Biocept, Inc., 5810 Nancy Ridge Drive Suite 150, San Diego, California 92121, USA
2Columbia University School of Engineering and Applied Science, Department of Chemical
Engineering, 

BIO Microfluidics_Biocept Paper.pdf

Detection of EpCAM-negative and cytokeratin-negative circulating tumor cells in peripheral blood 

Conclusions:
1.  Study demonstrates that the peripheral blood of cancer patients contain circulating tumor cells other than those normally detected with antibodies to EpCAM and cytokeratin.
2.  The use of CEE-Enhanced and antibody mixtures along with the traditional anti-EpCAM and anti-CK approach may lead to new insight into the diagnostic applications of CTCs.

Stephen D.Mikolajczyk,1 Lisa S.Millar,1 Pavel Tsinberg,1 StephenM. Coutts,1
MaryamZomorrodi,1 TamPham,2 Farideh Z. Bischoff,2 and Tony J. Pircher1
1Research and Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA
2Translational and Clinical Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA

http

http://www.hindawi.com/journals/jo/2011/252361/

http://www.hindawi.com/journals/jo/2011/252361 

Abstract & Posters

Detection of HER2 gene amplification in circulating tumor cells and disseminated tumor cells by fluorescence in situ hybridization using OncoCEETM - (SABC 2011)

Estrogen receptor and progesterone receptor immunocytochemistry staining in circulating tumor cells as compared to primary tumor or metastatic biopsy- (SABC 2011)

ER poster.pdf

Detection of HER2 status of circulating tumor cells and disseminated tumor cells using a microfluidic platform (cell enrichment and extraction technology [CEE^TM ]- (ASCO 2011)

Conclusion:
1.  The cell enrichment and extraction (CEE™) microfluidic technology provides a sensitive platform for evaluation of HER2 by FISH in intact CTCs and DTCs.  
2.  HER2+ CTCs and DTCs were encountered in patients with HER2+ as well as HER2- primary tumors.
3.  Discordant HER2 status was contributed mainly by HER2+ DTCs occurring in HER2- primary breast tumors.
4.  The clinical implications of evaluating HER2 in CTCs and DTCs in patients with invasive breast cancer needs further investigation. 

S Krishnamurthy¹, FZ Bischoff², K Wong²,S Mikolajczyk² , T Pham² , H.M. Kuerer, A Lodhi³ , A Bhattacaryya, C Hall³ ,  and A Lucci³
Universityof TexasMD Anderson Cancer Center Department ofPathology ¹, SurgicalOncology³, Biocept, Inc., San Diego CA²  

Biocept & MD Anderson CTC Poster ASCO 2011.pdf  

Redefining CTCs: Detection of additional circulating tumor cells using an antibody capture cocktail and HER2 FISH (ASCO 2011)

Conclusion:
1.  CEE™ technology provides a sensitive platform for enhanced capture, detection, and characterization of both CK+ and CK- CTCs.  
2.  This platform allows for evaluation of HER2 gene amplification status by FISH in intact CTCs within the microchannels at sensitivity and accuracy levels suitable for standardized CLIA-laboratory testing. 

Julie Ann Mayer, PhD, Tony J. Pircher, PhD, Stephen D. Mikolajczyk, Philip D Cotter, PhD, FACMG, Farideh Z. Bischoff, PhD,       Biocept Inc, San Diego, CA 92121  

Biocept Poster ASCO 2011_final.pdf

Redefining CTCs: Detection of cytokeratin-negative circulating tumor cells (CTCs) - AACR 2011

Conclusions:
1.  We have developed a robust method of detecting CTCs that can capture both epithelial and mesenchymal phenotypes.
2.  Our findings suggest current enrichment techniques may be missing an important population of CTCs.

ChadV. Pecot1, Farideh Bischoff2, Yvonne Lin3, Padmavathi Jaladurgam1, William Merritt4, Tony Pircher2, Steve Mikolajczyk2, Julie Mayer2, Karina Wong2, Tam Pham2, Justin Bottsford-Miller1, Rebecca Stone1, Joseph Celestino1, Alpa Nick1, Cathy Eng1, Anil Sood1

UT M.D. Anderson Cancer Ctr., Houston1, TX; Biocept Incorporated2, San Diego, CA; University of Southern California3, Los Angeles, CA; South Carolina Oncology4, Columbia, SC  

AACR_CTC Poster_2011_MD anderson_Pecot.pdf

FISH-based determination of HER2 status in circulating tumor cells isolated with the microfluidic CEE^TM  platform (AACR 2011)

Conclusions:
1.  CEE™ technology provides a sensitive platform for enhanced capture, detection, and characterization of both CK+ and CK- CTCs.  
2.  This platform allows for evaluation of HER2 gene amplification status by FISH in intact CTCs within the microchannels at sensitivity and accuracy levels suitable for standardized CLIA-laboratory testing.

Julie Ann Mayer, PhD, Tony J. Pircher, PhD, Stephen D. Mikolajczyk, Philip D Cotter, PhD, FACMG, Farideh Z. Bischoff, PhD,       Biocept Inc, San Diego, CA 92121 

 Mayer_HER2 by FISH and CTCs using microfluidic channel.pdf

Fluorescence labeling of cytokeratin-negative circulating tumor cells in peripheral blood: A new paradigm for the study of cancer progression (AACR 2011) 

Conclusions: The CEE-Enhanced staining technology enables the detection of CK-negative CTCs. This novel detection method, based on the in situ labeling of antibodies used for capture, greatly expands single and multi-antibody approaches to the study of rare circulating cells. Specifically, this allows the exploration and study of circulating cells not expressing CK such as highly de-differentiated tumor cells, stem cells or those tumor cells undergoing EMT. The clinical significance of these CK-negative CTCs is under investigation. 

Stephen D. Mikolajczyk, Lisa S. Millar, Maryam Zomorrodi, Farideh Z. Bischoff, Jayne Scoggin, Tam Pham, Karina Wong, Tony J. Pircher. Biocept, Inc., San Diego, CA  

AACR_2011_CTC paradigm shift_Mikolajczyk.pdf

Comparison of fluorescence in situ hybridization of estrogen receptor genetic locus with protein expression in invasive breast carcinoma  

CONCLUSION:
1. There is a significant heterogeneity between the gene amplification status and protein overexpression of ESR1.
2. The gene status of ESR1 ranges from negative, equivocal and amplified in both ER negative and ER immunopositive cases.
3. The significance of heterogeneity at the ESR1 gene locus in ascertaining the prognosis and predictive response to antiestrogen therapy needs further evaluation in larger prospective clinical trials. 

Julie A. Mayer¹, Tam Pham¹, Karina Wong¹, Anthony Lucci², Farideh Bischoff¹, Savitri Krishnamurthy². ¹Biocept Inc, San Diego, CA; ²The University of Texas MD Anderson Cancer Center, Houston, TX  

AACR_2011_ER FISH on CTC_Mayer.pdf

Detection of EpCAM-negative and Cytokeratin-negative Circulating Tumor Cells in Peripheral Blood (TriCon 2011)

Conclusions:
1.  Antibody mixtures improve the recovery of cancer cells, including CTCs not captured with anti-EpCAM alone
2.  CEE-Enhanced™ can be used to stain additional CTCs that do not stain with anti-cytokeratin
3.  CEE™ technology allows multiple screening and staining strategies for the analysis of CTCs

Stephen D. Mikolajczyk, Lisa S. Millar, Pavel Tsinberg, Maryam Zomorrodi, Jayne Scoggin, Tam Pham, Karina Wong, Farideh Z. Bischoff, Tony J. Pircher   Biocept Inc, San Diego, CA 92121

2011 Tri-Con CTC poster pdf.pdf 

Detection of circulating tumor cells and HER2 gene amplification status in breast cancer using a novel microfluidic platform (Cell Enrichment and Extraction Technology, CEE^TM

Conclusions:
1. The cell enrichment and extraction microfluidic technology (CEE) provides a sensitive platform for enhanced detection and characterization of antibody cytokeratin positive and negative CTCs.
2. This platform allows evaluation of HER2 gene amplification status by fluorescence in situ hybridization (FISH) in intact CTCs within the microchannels.
3. The utility of this platform for phenotypic and genotypic characterization of CTCs in breast cancer needs to be tested in larger clinical trials.

Savitri Krishnamurthy¹, Farideh Z. Bischoff², Steve Mikolajczyk²,and Anthony Lucci³
Universityof TexasMD Anderson Cancer Center Department ofPathology ¹, SurgicalOncology³, Biocept, Inc., San Diego CA² 

 San Antonio Poster For Dec 2010.pdf

Efficient Capture of Suspended Tumor Cells with a Novel Immunocytochemical Microfluidic Device

Conclusions:
1. Extremely rare cells can be separated with high efficiency and high purity for in situ inspection.
2. Higher antigen densities yield higher capture.  Middle flow rates yield constant k values, avoiding streamline effect.

Mary Dickson†, ‡, Pavel Tsinberg†, Zhongliang Tang†, Timothy Wilson†, Edward Leonard‡
†Biocept, Inc., San Diego, CA
‡Department of Chemical Engineering, Columbia University, New York, NY

Dickson Poster.pdf

CEE™ CELL ENRICHMENT AND EXTRACTION TECHNOLOGY- Enabling Early, Non-invasive Isolation of Rare Cells for Diagnostics

CONCLUSIONS: Standard marker and FISH analysis of the cells can be completed within the device for highly accurate diagnosis. 

CEE white paper 0106.pdf  

Isolation of Epithelial and Mesenchymal (EMT) Tumor Cell Lines From Blood Using Multiple Capture Antibodies in a Microfluidic Cell Capture System

Stephen D. Mikolajczyk1, Tony J. Pircher1, Maryam Zomorrodi1, Farideh Z. Bischoff1, Michael B. Williams2 , and Ashish M. Kamat2 

1. Biocept, Inc., San Diego, CA
2. MD Anderson Cancer Center, Dept of Urology, Houston, TX, Department of Urology, Houston, TX 

NCI CTC Conf 0909[1].pdf

Mouse Models for MEMS (Micro-Electro-Mechanical System) Based Biomarker Discovery: A Novel CTC Mouse-Trap

Farideh Z. Bischoff1, Pavel Tsinberg1, William M. Merritt2, Yvonne G. Lin2, Anil K. Sood2

1. Biocept, Inc., San Diego, CA
2. MD Anderson Cancer Center, Dept of Gynecologic Oncology, Houston, TX, Department of Urology, Houston, TX 

Oncology Biomarkers 2008[1].pdf

Recovery of Circulating Tumor Cells in Urothelial Carcinoma Cell Lines Utilizing a Novel Antibody Based Microfluidic Cell Capture Technique

Michael Williams1, Tony Pircher2, Maryam Zomorrodi2, Steve Mikolajczyk2, Ashish Kamat1

1Urology, MD Anderson Cancer Center
2Biocept 

AUA-2010 technique submission.pdf