Affiblot: a dot blot-based screening device for selection of reliable antibodies

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081715%3A_____%2F21%3A00545112" target="_blank" >RIV/68081715:_____/21:00545112 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216275:25310/21:39918249 RIV/00216208:11160/21:10431693

Výsledek na webu

<a href="http://hdl.handle.net/11104/0321871" target="_blank" >http://hdl.handle.net/11104/0321871</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1039/d1ay00955a" target="_blank" >10.1039/d1ay00955a</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Affiblot: a dot blot-based screening device for selection of reliable antibodies

Popis výsledku v původním jazyce

The key factor in the development of antibody-based assays is to find an antibody that has an appropriate affinity, high specificity, and low cross-reactivity. However, this task is not easy to carry out since the research antibodies on the market may suffer from low specificity and reproducibility. Here, we report on a palm-sized dot blot-based device, called the affiblot, that has a specially designed lid that allows simultaneous semi-quantitative comparison of up to five antibodies from different suppliers regarding their affinity/avidity, cross-reactivity, and batch-to-batch reliability. The only required peripheral equipment is a vacuum pump, a camera, and densitometry software. The affiblot device was tested for its functionality and its measurements were compared against those obtained by standard dot blot and ELISA. The benefit over these methods, when various antibodies are evaluated, is in its simplicity. It allows easy antigen deposition, fast application and the discarding of the solutions, a compact undivided membrane, and therefore significant decrease of labor. The device was tested with specific anti-ApoE, anti-EpCAM, anti-Salmonella, anti-E. coli, and anti-Listeria antibodies from different suppliers. Their properties were compared for their ability to interact specifically with antigen and/or non-target structures and the best-suited antibody for the intended application was identified.

Název v anglickém jazyce

Affiblot: a dot blot-based screening device for selection of reliable antibodies

Popis výsledku anglicky

The key factor in the development of antibody-based assays is to find an antibody that has an appropriate affinity, high specificity, and low cross-reactivity. However, this task is not easy to carry out since the research antibodies on the market may suffer from low specificity and reproducibility. Here, we report on a palm-sized dot blot-based device, called the affiblot, that has a specially designed lid that allows simultaneous semi-quantitative comparison of up to five antibodies from different suppliers regarding their affinity/avidity, cross-reactivity, and batch-to-batch reliability. The only required peripheral equipment is a vacuum pump, a camera, and densitometry software. The affiblot device was tested for its functionality and its measurements were compared against those obtained by standard dot blot and ELISA. The benefit over these methods, when various antibodies are evaluated, is in its simplicity. It allows easy antigen deposition, fast application and the discarding of the solutions, a compact undivided membrane, and therefore significant decrease of labor. The device was tested with specific anti-ApoE, anti-EpCAM, anti-Salmonella, anti-E. coli, and anti-Listeria antibodies from different suppliers. Their properties were compared for their ability to interact specifically with antigen and/or non-target structures and the best-suited antibody for the intended application was identified.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/EF17_048%2F0007421" target="_blank" >EF17_048/0007421: Posilování mezioborové spolupráce ve výzkumu nanomateriálů a při studiu jejich účinků na živé organismy</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Analytical Methods: advancing methods and applications

ISSN

1759-9660

e-ISSN

1759-9679

Svazek periodika

13

Číslo periodika v rámci svazku

35

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

11

Strana od-do

3874-3884

Kód UT WoS článku

000687267500001

EID výsledku v databázi Scopus

2-s2.0-85115395353