definition of Precipitation assays

performed in semi-solid media such as agar or agarose where antibodies and antigens can diffuse toward one another and form a visible line of precipitation

difference between precipitation and agglutination

Agglutination is the process of clumping of antigens with their respective antibodies. Precipitation is a process where soluble antigens bind with their specific antibody at an optimum temperature and pH, resulting in the formation of an insoluble precipitate

most abundant type of antibody in the body, comprising most of the immunoglobulin in secretions and a significant amount of circulating immunoglobulin -> which immunoglobulin?

igA

purpose of igA

protect the mucosal tissues from microbial invasion and maintain immune homeostasis with the microbiota

usually of higher affinity and are found in blood and in extracellular fluid -> which immunoglobulin?

igG

purpose of igG

neutralize toxins, viruses, and bacteria, opsonize them for phagocytosis, and activate the complement system

why measurement of IgM levels is very important

evaluate the progression of viral and bacterial infections

first antibody to be synthesized by activated-B cells in a primary immune response against a certain antigen -> which immunoglobulin

igM

most useful for determining recent infection -> which immunoglobulin

igM

usually detectable for longer periods following an infection -> which ig

igG

usually becomes undetectable weeks to months following infection -> which ig

igM

regulatory role, acting as an anti-inflammatory, preventing allergic reactions, and protecting the body from autoimmune diseases and severe infections -> which ig

igD

found in serum in much smaller quantities than other Ig classes -> which ig(s)

igD and igE

primarily defends against parasitic invasion and is responsible for allergic reactions -> which ig

igE

receptor for antigens found mostly on mature B-lymphocytes -> which ig

igD

what are the types of ig

igA, igG, igM, igD, igE

possess heavy chains known as γ-chains -> which ig

igG

how many chains and what arrangements immunoglobulins have?

two light chains and two heavy chains in a light-heavy-heavy-light structure arrangement

has a highest molecular weight -> which ig

igM

the most abundant antibody present in newborns, also the only immunoglobulin that crosses the placenta -> which ig

igG

the major antibody in secretions found in saliva, tears, colostrum, intestinal, genital tract, and respiratory secretions -> which ig

igA

most common immunodeficiency, characterized by recurrent infections that affect the respiratory, digestive, and genitourinary systems?

Selective IgA deficiency

protein in each zone alpha1, alpha2, beta, gamma

later (textbook protein chapter)

each result of immunosubtraction correspond with what disease state

later (part 3 and 4 serology)

first developed IA?

radioimmunoassay (RIA)

Five types of immunoassay

enzyme immunoassay (EIA), radioimmunoassay (RIA), fluoroimmunoassay (FIA), chemiluminescent immunoassay (CLIA) and counting immunoassay (CIA)

have excess antibody binding sites and produce a signal directly proportional to the amount of analyte in the sample -> which type of IA

Non-competitive immunoassay

limited the number of antigen binding sites, inversely proportional to the amount of analyte of interest -> which type of IA

Competitive immunoassay

The antigen is detected by a primary antibody, conjugated with a label for detection (enzyme, fluorophore, etc.) -> which type of IA

Direct Immunoassay

An unconjugated primary antibody binds to the antigen. A secondary antibody conjugated with a detection label binds to the primary antibody -> which type of IA

Indirect Immunoassay

Amplified signal due to multiple secondaries binding to primary -> which type of IA

Indirect Immunoassay

use an antibody linked to an enzyme to detect and measure antigens or antibodies -> which type of IA

Enzyme immunoassays (EIA)

most common and popular quantitative immunoassay technology. This type of immunoassay employs microplates to immobilize a target antigen or antibody, bind with a targeted analyte, and measure an emitted signal -> which type of IA

ELISA

single, enzyme-linked antibody directly binds with an antigen; useful when analyzing immune responses to antigens -> which IA

Direct ELISA

enzyme-linked secondary antibody binds with a primary antibody bound to the antigen; useful when determining total antibody concentration -> which IA

Indirect ELISA

pros and cons of direct ELISA

This is ELISA in its simplest form but suffers from low flexibility and potentially high background signal. Direct ELISAs are useful when analyzing immune responses to antigens.

pros and cons of indirect ELISA

The secondary antibodies make this ELISA highly versatile, sensitive, and cost-effective. However, the secondary presents the potential for cross-reactivity and adds complexity to the protocol. Indirect ELISAs are useful when determining total antibody concentration.

An antigen binds to a primary antibody attached to the plate. A secondary enzyme-linked antibody then attaches to the antigen -> which IA

Sandwich ELISA

pros and cons of sandwich ELISA

highly flexible, sensitive, and specific but requires large antigens of interest to accommodate multiple antibody bindings. Sandwich ELISAs are particularly useful when analyzing complex samples, since the antigen doesn’t need to undergo purification before measurement

The sample antibodies compete with enzyme-linked antibodies to bind with the antigen -> which IA

Competitive ELISA

Fill in the blank with positive/negative:

The capillary tube end dipped in the buffer with the anode behaves as a ___ side and the end with the cathode behaves as the ___ side of the electrophoretic system. Capillary tube walls have a ___ charge and ___ ions in the buffer get attracted to and neutralize it

positive, negative, negative, positive

capillary electrophoresis testing what

protein abnormalities testing

Capillary Electrophoresis, all proteins migrate toward cathode/anode?

cathode

main sample type for Capillary Electrophoresis

urine