rabbit anti-her2 monoclonal antibody (clone ep3)  (ZSGB Biotech)

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Defining the optimized cutoff for 4B5 using receiver operating characteristic (ROC) analysis based on the standard protocol of FISH/IHC analyses. (A) ROC analysis of QDB results. (B) The distribution of HER2 levels within each IHC category. All the FFPE specimens in cohort 1 were categorized as Her2+ and Her2− based on combined IHC and FISH analyses. FISH results superseded the IHC results whenever a conflict occurred. Her2 levels measured by the QDB method using the 4B5 antibody were plotted against categorized Her2 results for ROC analysis ( n = 319) in (A) . In (B) , Her2 levels measured with QDB were plotted against each IHC categorized scores. Log scale was used to illustrate the performance of the proposed cutoff at 0.399 nmol/g. For specimens with undetectable Her2 levels, a value at 0.001 nmol/g was arbitrarily entered to avoid missing any specimen in the graph.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Absolutely quantitative analysis of Her2 expressions in cohort 2. (A) The distribution of HER2 levels measured with the QDB method using 4B5 and EP3 antibodies respectively in cohort 2. The results were expressed as mean ± SEM. (B) Correlation analysis of the Her2 levels in cohort 2 measured with the QDB method using 4B5 and EP3 antibodies, respectively. Pearson’s correlation analysis was performed with r = 0.981, p < 0.0001. (C) Correlation of HER2 levels measured with QDB using the 4B5 antibody with those from IHC analysis. The results were analyzed using Spearman’s correlation analysis with ρ = 0.545, p < 0.0001.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Overall distribution of absolutely quantitated HER2 levels in the merged cohort within each IHC category. The Her2 protein levels in both cohort 1 and cohort 2 were measured with the QDB method using EP3 and 4B5 antibodies, respectively, and plotted together against their respective IHC scores. (A) In QDB analysis with the 4B5 antibody, Her2 levels ranged from 0 to 65.05 nmol/g, with a mean of 2.361 ± 0.290 and a median of 0.108, n = 577. (B) For those with the EP3 antibody, its levels ranged from 0 to 31.31 nmol/g, with a mean of 1.419 ± 0.166 and a median of 0.048, n = 578.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Comparison of dichotomized QDB results measured with 4B5 and EP3 using their respective cutoffs in the merged cohort (cohorts 1 and 2; n = 578).

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Comparison

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Sensitivity and specificity of Her2 levels from the QDB method using 4B5 and EP3 antibodies, respectively, vs. IHC ( n = 578) using FISH results as the gold standard.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Evaluation of the distribution of breast cancer specimens with no Her2 expression (Her2-0) from those with any Her2 expression (Her2-E) in each IHC category based on the limit of detection (LOD) of the QDB method using 4B5 antibodies.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: The distribution of specimens with (Her2-E) and without Her2 expression (Her2-0) stratified by limit of detection (LOD) of QDB analysis using either EP3 or 4B5 antibodies. The LODs of QDB analysis with both 4B5 and EP3 antibodies were at 0.09 nmol/g. Specimens with Her2 levels above 0.09 nmol/g with both antibodies were in gray color while those below 0.09 nmol/g were in the white circle. Specimens with Her2 levels above 0.09 nmol/g only when measured using the 4B5 antibody were in brown color while those only with the EP3 antibody were in blue color.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: The clinicopathological characteristics of the patients.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques:

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Correlation of HER2 levels measured with 4B5 and EP3 antibodies. A total of 332 breast cancer FFPE tissues in 2 × 5 µm slices were provided by a local hospital. MCF-7 and BT474 cell lysates were used as internal controls. FFPE tissue lysates (about 0.5 µg/unit) and cell lysates (about 0.3 µg/unit) were applied onto the QDB plates at 2 µl/unit in triplicate for the QDB measurements with clone EP3 and 4B5 respectively. A set of serially diluted HER2 recombinant protein were included in each plate to develop plate-specific standard curve. All results were averaged from three independent experiments, with each sample in triplicate. The correlation of HER2 levels measured with 4B5 and EP3 was analyzed with Pearson’s correlation coefficient analysis using Graphpad software, r = 0.963, p < 0.0001.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Recombinant, Software

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Distribution of all 332 samples. HER2 levels in all 332 breast cancer FFPE sample lysates were measured with the QDB method using EP3 antibody. The lysates were diluted to about 0.25 µg/µl, and then 2 µl lysate was used for each sample. ( a ) the distribution of HER2 levels among 332 samples. HER2 levels ranged from 0 (chemiluminescence readings less than two fold over the background) to 31.310 nmol/g. ( b ) All samples were grouped by their IHC scores provided by local hospital. The distributions of HER2 levels in each IHC group were recorded as following: 0, 0–0.205 nmol/g, mean = 0.045 ± 0.006 nmol/g, n = 77; 1 +, 0 –0.410 nmol/g, mean = 0.049 ± 0.008 nmol/g, n = 65; 2+ , 0–7.250 nmol/g, mean = 0.537 ± 0.122 nmol/g, n = 108; and 3+ , 0.329–31.310 nmol/g, mean = 7.120 ± 0.773 nmol/g, n = 82. The intra- and inter-CV were 8.98% and 9.89% respectively.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques:

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Evaluation of QDB results with those of IHC and FISH analyses using ROC analysis. Samples were separated into negative (HER2−) and positive (HER2+) groups based on the recommendations from ASCO/CAP. In ( a ), samples were grouped based on their IHC scores, with 142 samples in the negative group (IHC 0 and 1+), and 82 samples in the positive group (IHC 3+). Absolute HER2 levels from these samples were used for ROC analysis with Graphpad Prism7.0 software. The ROC curve of QDB results was obtained with area under the Curve (AUC) at 0.9998 ± 0.0002; 95% CI 0.9994–1; p < 0.0001. In ( b ), samples were grouped based on both IHC and FISH results, with 214 samples as negative (HER2−) group and 105 samples as positive (HER2+) group. 6 equivocal cases and 7 cases with missing FISH results were excluded in the analysis. Absolute HER2 levels from these samples were used for ROC analysis with Graphpad Prism7.0 software. The area under the curve (AUC) was at 0.9942 ± 0.0031, with 95% CI at 0.9881–1; p < 0.0001. ( c ) The samples were grouped by IHC scores, and the suggested cutoff values from ROC analyses in ( a ) at 0.267 nmol/g (solid line), and in ( b ) at 0.261 nmol/g (dashed line) were shown to demonstrate the effectiveness of these cutoff values to separate samples from HER2+ to HER2− groups. HER2 levels were plotted in log scale to better demonstrate the distribution of QDB results among these samples. For those samples with undetectable HER2 level, a value of 0.001 nmol/g was arbitrarily entered to avoid omitting any sample in the log scale graph.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Software

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Assessing HER2 levels by histologic grade. The FFPE specimens (300 out of 332) were grouped according to their Nottingham histologic scores into Grades I, II, and III. The HER2 levels of each grade were used for column statistics analysis with Graphpad Prism7.0 software. The mean ± SD of the HER2 levels were 0.791 ± 0.555 nmol/g for Grade I (n = 36), 1.554 ± 0.330 nmol/g for Grade II (n = 145), and 3.271 ± 0.535 nmol/g for Grade III (n = 119). The statistical difference was assessed with two-tailed Student’s t-test, with p < 0.05 between Grades I and III, and p = 0.005 between Grades II and III. There was no statistical difference between Grade I and Grade II samples.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Software, Two Tailed Test