Immunology and Serology LAB
antigen
Immunology study of hosts reaction when foreign
substances introduce in the body
- Antigen foreign substance that elicit the
production of antibody
- Antibody specific glycoprotein produced in
response to antigenic challenge
Serology detection/measurement of humoral immune
response
- Antibody
- Serum
Plasma Vs. Serum
Plasma is the liquid, dell-free part of blood that
has been treated with anticoagulants. (anticoagulated)
Serum is the liquid part of blood after
coagulation, therefore devoid of clotting factors as
fibrinogen. (clotted)
Serum = Plasma fibrinogen
3 Distinct Phases of Ag and Ab Reactions
- Primary phenomenon sensitization
- Secondary Phenomenon lattice formation
- Tertiary phenomenon detected by affect on
tissues or cells
Primary Phenomenon
Sensitization binding of antibody to antigen
not visible
Secondary Phenomenon
- Lattice formation
- The Fab portion of the Ig molecule attaches to
antigens on 2 adjacent cells visible results in
agglutination
- If both antigen and antibody are SOLUBLE,
reaction will become visible over time, i.e.
Precipitation
Tertiary Phenomenon
Reaction not visible, detected by affect of
reaction on tissues or cells = phagocytosis
Precipitation soluble antibody reacts with soluble
Precipitation Curve
- Prozone antibody in excess false negative
- Postzone antigen in excess false negative
- Zone of equivalence Ag and Ab present in
optimal proportion and give visible reaction
Agglutination particulate antigens bound together by
antibody
- Direct Agglutination like in blood typing
- Passive Agglutination employs particles that
are coated with antigens, i.e. RBCs, polystyrene
latex, bentonite or charcoal
- Reverse Passive Agglutination Ab attached to
carrier particle instead of Ag
- Agglutination Inhibition competition between
particulate and soluble antigens for limited
antibody binding site
- Coagglutination bacteria is the inert particles
Rapid Flow Chromatographic Immunoassay
Rapid test cassette kit
Blood Specimen Preparation
- Either serum or plasma
- After the blood is collected, allowed to clot for
30 minutes
- Centrifuge then transfer the serum in a labeled
test tube
- Conduct the testing immediately
- If cannot be performed, it should be frozen at
-20C
Clinical Chemistry LEC Cellular Components (45% total Blood volume)
- Buffy coat
Clinical Chemistry o WBC 7,000-9,000 per mm3 of blood
- Basic science that utilizes the specialty of o Platelets 250,000 per mm3 of blood
chemistry to study human beings - RBC about 5,000,000 per mm3 of blood
- Applied science where analyses are performed
on body fluids or tissues for diagnosis and Blood Proteins
treatment of disease - Albumin, Globulin and Fibrinogen
Nutrients
Different analytes: - Amino acids, Sugar, Lipids
- Glucose Hormones
- Non-Protein Nitrogen Compound - Erythropoietin, Insulin
o BUN Electrolytes
o Creatinine derived from creatine - Calcium, Sodium, Potassium
o Uric Acid
o Ammonia
- Lipids
o HDL
o LDL
o Total Cholesterol
o Triglycerides
- Proteins
o Total Protein Biomarkers analytes that determine if patient is at risk
o Albumin Glycoprotein Hemoglobin uses whole blood
o Globulin
- Amino acids Composition of Cells
- Plasma/Cell Membrane separate the cells
Malfunction: from the surrounding environment
- Trauma or by invasive agents o Primary structure: double layer of
- Genetic deficiency of vital enzyme phospholipid molecules
- Insufficient supply of essential nutrients o Membrane proteins controls what
- Insufficient blood and oxygen supply moves through the membrane by
- Malignancy means of membrane proteins
- Accumulation of waste products embedded in the phospholipid layer
- Defect in the cellular recognition of signals o Membrane proteins + carbohydrate
forms glycoproteins that act as
Role of Clinical Chemistry Laboratory: identification markers
- To measure the concentration of particular o Receptors react to specific chemicals
constituents (or analytes) in body fluids or - Cytoplasm thick gel-like substance inside the
tissues cell that is composed of numerous organelles
suspended in watery cytosol (intracellular fluid)
Plasma (55% total blood volume) - Nucleus large membranous structure near the
- 91% water center of the cell
- 7% blood proteins o Control center of the cell
- 2% Nutrients, Hormones, Electrolytes
- Endoplasmic Reticulum membranous-walled
canals and flat, curing sacs arranged in parallel
tows throughout the cytoplasm
o Rough ER has ribosomes
Protein synthesis
Intracellular transportation
o Smooth ER no ribosomes
Lipid and carbohydrate
synthesis
Creates membranes for use
- Ribosomes non membranous structure made
of two pieces, a large subunit and a small
subunit, each subunit is composed of rRNA
o Ribosomes in the ER make proteins for
export or to be embedded in the
plasma membrane
o Free ribosomes make proteins for the
cells domestic use
- Golgi Apparatus consists of cisternae stacked
on one another and located near the nucleus
o Processes/packages protein molecules
from the ER
o Processed proteins leave the final
cistern in a vesicle; contents may then
be secreted outside the cell
- Lysosomes microscopic membranous sacs
that have pinched off the Golgi Apparatus
o Cells own digestive system
o Apoptosis programmed cell death
- Peroxisomes contains enzymes that detoxify
harmful substances that enter cells
o Often seen in kidney and liver cells
- Mitochondria walls composed of inner and
outer membranes separated by fluid
o Powerplants of cells
o Mitochondrial enzymes catalyze series
of oxidation-reactions that provide
about 95% of the cells energy supply
- Cytoskeleton internal supporting framework
made up of rigid, rod-like pieces that provide
support and allow movement
- Cell fibers intricately arranged fibers of
varying lengths that form a three-dimensional,
irregularly shaped lattic
o Supports the ER, mitochondria, free
ribosomes
- Microfilaments cellular muscles; can slide past
each other causing shortening of the cell
- Intermediate filaments supporting framework
in many types of cells
- Microtubules made up of protein subunits
arranged in a spiral fashion;
o moves around the cell
- Centrosome an area of the cytoplasm near
the nucleus that coordinates the building and
breaking of microtubules of the cells
o Non-membranous
o Microtubule-organizing center
o Plays important role during cell division
- Centriole identifies the general location of the
centrosome
- Cell extensions projections that extend the
plasma membrane outward to form tiny,
fingerlike processes
o Microvilli absorption
Surface area
Epithelial cells in intestine
o Cilia and Flagella microtubules at their
core; sperm cells
- Nucleus contains DNA
o Nuclear envelope composed of two
membranes each with essentially the
same molecular structure as plasma
membranes
o Nucleoplasm
o Nuclear pores holes in the nuclear
envelope
- DNA determines bothe structure and
functions of cells and heredity;
appears as:
o Chromatin threads/granules in non-
dividing cells
o chromosomes