Question

Which techniques could be used to identify antibodies produced by a patient?

Answer 1

Explanation:

For the detection of the antibody, several methods can be used:

• By Peroxidase

Peroxidase is an enzyme that uses H2O2 to oxidize its substrates. These substrates are designed in such a way that their oxidation can be detected by light release (luminol), by the formation of a colored precipitate (DAB) or by the transformation of a colorless compound into a colorful compound, can be determined quantitatively by spectroscopy. Luminol, in the presence of H2O2 and peroxidase emits light, which can be captured by X-ray films, being mainly used in immunodetection. Diaminobanzidine (DAB), in the presence of H2O2 and peroxidase, forms a colored polymer brown, precipitating in the place where the peroxidase is found, being mainly used in immunocytochemistry studies. There is a wide variety of compounds that can be used as chromophores, providing the most varied colors, usable in ELISA technique, with ABTS (2, 2'-azino-di (3-ethylbenzthiazoline-6-sulfonate) being an example of dye that turns blue-green in the presence of peroxidase.

• By Alkaline phosphatase

Alkaline phosphatase is an enzyme that hydrolyzes phosphate bonds. In this way, chromophores are compounds that have phosphate groups, and that with the loss of this group phosphates acquire color, as is the case with p-nitrophenyl phosphate, which turns yellow when phosphate group.

• By Fluorophore

Fluorophore is a compound that emits light of a certain wavelength (λ (coloring) when excited by a light from another λ. Fluorophores with different characteristics make it possible to mark different antigens with different colors in the same tissue or cell, allowing simultaneous observation oftwo or more components.

• By Metals with high density

The main heavy metal conjugated to antibodies is gold. This allows you to locate antigens through electron microscopy, allowing very specific cytological studies