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