|
| Status |
Public on Apr 15, 2023 |
| Title |
INF 01-PI |
| Sample type |
protein |
| |
|
| Source name |
Serum-PI
|
| Organism |
Homo sapiens |
| Characteristics |
subclass: Infant
tissue: Blood
treatment: PI (pre-immune treatment)
|
| Extracted molecule |
protein |
| Extraction protocol |
Human pool sera (25 subjects) from NCT00937521
|
| Label |
AlexaFluor 647-conjugated anti-human IgG secondary antibody
|
| Label protocol |
The secondary antibody used for arrays was from Jackson Immunoresearch, (872 W Baltimore Pike, West Grove, PA 19390 USA). It was conjugated to the Alexafluor dye (Alexa 647) by the company.
|
| |
|
| Hybridization protocol |
Protein arrays were first blocked 60 min in Blocking Buffer (Block it, Arrayit). The array was then incubated 1h a room temperature (RT) with human sera diluted 1:1000 for anti-human IgG-specific analysis. After two washing with 0.1% Tween in PBS buffer (TPBS), arrays were incubated with an AlexaFluor 647-conjugated IgG anti-human (1:800 – Jackson Immunoresearch) at RT for 1 h to detect interactions. Slides were washed again as before and washed once in PBS, then rinsed in sterile water and dried.
|
| Scan protocol |
Fluorescence signals were detected by using a Powerscanner (Tecan Trading ) and the 16-bit images were generated with Powerscanner software at 10 µm per pixel resolution.
|
| Description |
NCT00937521
FS00009245_Area1-INF 01-PI
|
| Data processing |
Images were processed using ImaGene 9.0 software (Biodiscovery Inc, CA, USA). Elaboration and analysis of image raw fluorescence Intensity (FI) data was performed using in-house developed R scripts. For each protein the fluorescence intensity data of the replicates was subtracted by local background values surrounding each spot and averaged, hence resulting in a single data point per sample named Mean Fluorescence Intensity (MFI).
Mean fluorescence intensity (MFI)
|
| |
|
| Submission date |
Feb 02, 2023 |
| Last update date |
Apr 16, 2023 |
| Contact name |
Erika Bartolini |
| Organization name |
GSK Vaccines
|
| Department |
AIMB
|
| Lab |
Molecular Biology
|
| Street address |
Via Fiorentina 1
|
| City |
Siena |
| State/province |
Italy |
| ZIP/Postal code |
53100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL33085 |
| Series (2) |
| GSE224340 |
OpcA and PorB are novel bactericidal antigens of the 4CMenB vaccine in mice and humans |
| GSE224348 |
OpcA and PorB are novel bactericidal antigens of the 4CMenB vaccine in mice and humans |
|
