PMID- 32719400
OWN - NLM
STAT- MEDLINE
DCOM- 20201209
LR  - 20210727
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 10
IP  - 1
DP  - 2020 Jul 27
TI  - Developing a routine lab test for absolute quantification of HER2 in FFPE breast 
      cancer tissues using Quantitative Dot Blot (QDB) method.
PG  - 12502
LID - 10.1038/s41598-020-69471-4 [doi]
LID - 12502
AB  - Developing immunoassay for absolute quantitation of protein biomarkers in 
      Formalin Fixed Paraffin Embedded (FFPE) samples promises improved objectivity, 
      consistency and accuracy in daily clinical practice. The feasibility of 
      Quantitative Dot Blot (QDB) method for this purpose was explored in this study. 
      We were able to measure HER2 protein levels using 0.5 microg/sample total protein 
      lysate extracted from 2 x 5 microm FFPE slices absolutely and quantitatively using 
      QDB method in 332 breast cancer FFPE samples. HER2 levels measured using two 
      clinically validated antibodies for immunohistochemistry respectively were highly 
      correlated (r = 0.963). We also achieved area under the curve (AUC) at 
      0.9998 +/- 0.0002 (p < 0.0001, n = 224) with IHC analysis, and 0.9942 +/- 0.0031 
      (p < 0.0001, n = 319) with combined results from IHC and Fluorescence in situ 
      hybridization (FISH) analyses when analyzed with Receiver Operative 
      Characteristics analysis (ROC) respectively. When the results were converted 
      dichotomously with optimized cutoffs from ROC analyses, we achieved 99.5% 
      concordance with IHC; and 96.9% with combined results from both IHC and FISH 
      analyses. Therefore, we were able to demonstrate QDB method as the first 
      immunoassay platform for absolute quantitation of protein biomarkers in FFPE 
      samples to meet the need of daily clinical practice, especially for local 
      laboratories or laboratories in developing countries.
FAU - Yu, Guohua
AU  - Yu G
AD  - Lab of Molecular Pathology, Dept of Pathology, Affiliated Yuhuangding Hospital of 
      Qingdao University, Yantai, Shandong, People's Republic of China.
FAU - Zhang, Wenfeng
AU  - Zhang W
AD  - Yantai Quanticision Diagnostics, Inc., a Division of Quanticision Diagnostics, 
      Inc., of USA, Yantai, Shandong, People's Republic of China.
FAU - Zhang, Yunyun
AU  - Zhang Y
AD  - Yantai Quanticision Diagnostics, Inc., a Division of Quanticision Diagnostics, 
      Inc., of USA, Yantai, Shandong, People's Republic of China.
FAU - Lv, Jiahong
AU  - Lv J
AD  - Yantai Quanticision Diagnostics, Inc., a Division of Quanticision Diagnostics, 
      Inc., of USA, Yantai, Shandong, People's Republic of China.
FAU - Wu, Shishou
AU  - Wu S
AD  - Lab of Molecular Pathology, Dept of Pathology, Affiliated Yuhuangding Hospital of 
      Qingdao University, Yantai, Shandong, People's Republic of China.
FAU - Sui, Xiaolong
AU  - Sui X
AD  - Lab of Molecular Pathology, Dept of Pathology, Affiliated Yuhuangding Hospital of 
      Qingdao University, Yantai, Shandong, People's Republic of China.
FAU - Zhang, Jiandi
AU  - Zhang J
AD  - Yantai Quanticision Diagnostics, Inc., a Division of Quanticision Diagnostics, 
      Inc., of USA, Yantai, Shandong, People's Republic of China. 
      jiandi.zhang@outlook.com.
AD  - Center for Precision Medicine, Binzhou Medical University, Yantai, Shandong, 
      People's Republic of China. jiandi.zhang@outlook.com.
AD  - Quanticision Diagnostics, Inc., 400 Park Offices Dr., Research Triangle Park, NC, 
      27709, USA. jiandi.zhang@outlook.com.
FAU - Tang, Fangrong
AU  - Tang F
AD  - Yantai Quanticision Diagnostics, Inc., a Division of Quanticision Diagnostics, 
      Inc., of USA, Yantai, Shandong, People's Republic of China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200727
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Neoplasm Proteins)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Breast Neoplasms/*metabolism/pathology
MH  - Cell Line, Tumor
MH  - Female
MH  - Humans
MH  - *Immunoblotting
MH  - Middle Aged
MH  - Neoplasm Grading
MH  - Neoplasm Proteins/metabolism
MH  - *Paraffin Embedding
MH  - Receptor, ErbB-2/*metabolism
MH  - Reproducibility of Results
MH  - *Tissue Fixation
PMC - PMC7385113
COIS- WZ, FT, YZ, JL & JZ are employees of Yantai Quanticision Diagnostics, Inc., a 
      division of Quanticision Diagnostics, Inc., who own or has filed patent 
      applications for QDB plate, QDB method, & QDB application in IHC analysis. GY, 
      SW, XS declared no competing interests.
EDAT- 2020/07/29 06:00
MHDA- 2020/12/15 06:00
PMCR- 2020/07/27
CRDT- 2020/07/29 06:00
PHST- 2019/11/14 00:00 [received]
PHST- 2020/07/02 00:00 [accepted]
PHST- 2020/07/29 06:00 [entrez]
PHST- 2020/07/29 06:00 [pubmed]
PHST- 2020/12/15 06:00 [medline]
PHST- 2020/07/27 00:00 [pmc-release]
AID - 10.1038/s41598-020-69471-4 [pii]
AID - 69471 [pii]
AID - 10.1038/s41598-020-69471-4 [doi]
PST - epublish
SO  - Sci Rep. 2020 Jul 27;10(1):12502. doi: 10.1038/s41598-020-69471-4.