(M) IS Lab L1

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Cellulose Acetate

Standard, inexpensive, but has larger pores and lower resolution.

Agarose Gel

Finer pore structure, better resolution, and preferred for clinical use.

albumin > alpha-1 > alpha-2 > beta > gamma

Major Serum Proteins ordered from fastest to slowest migration: __________ > __________ > __________ > __________ > __________

Prealbumin

Slightly faster than albumin.

Albumin

Most abundant major protein and the fastest.

Ponceau S

__________ is a standard red dye for proteins on membranes

Sulfosalicylic acid

SPE staining: __________ and Ponceau S are used.

Normal Peak

High spike in albumin, with a small, flat, smooth bump in the gamma region.

Polyclonal

Broad, diffuse increase in the gamma region representing a wide variety of antibodies, typical of chronic infection or inflammation.

Monoclonal

A sharp, narrow spike (M-spike) in the gamma or beta region. Caused by the uncontrolled proliferation of a single plasma cell clone (e.g., Multiple Myeloma).

Multiple Myeloma

A malignancy of plasma cells usually producing excess IgG or IgA and light chains.

Waldenström Macroglobulinemia

A B-cell lymphoma producing excess, heavy IgM, leading to hyperviscosity syndrome.

AL Amyloidosis

Deposition of insoluble monoclonal light chain fibrils in tissues.

Immunofixation Electrophoresis (IFE)

While SPE identifies that a monoclonal spike exists, __________ tells you exactly what type of immunoglobulin is causing the spike.

Serum Protein Electrophoresis (SPE)

__________ separates serum proteins according to their electrophoretic mobility.

Capillary Zone Electrophoresis (CZE)

__________ is a widely used form utilizing very small fused-silica capillary tubes.

Chromatography

__________ separates substrates according to their distribution and different migration-separation characteristics between a mobile phase and a stationary phase.

Gas Chromatography

Mobile phase: Inert gas (e.g., Helium, Nitrogen)

Stationary phase: Liquid or solid coated on a column wall

Liquid Chromatography

Mobile phase: Liquid solvent

Stationary phase: Solid particles packed in a column

Column Chromatography

Mobile phase: Liquid moving through the column

Stationary phase: Solid beads inside the glass/plastic column

Flat Methods: Thin-Layer (TLC)

Mobile phase: Liquid solvent

Stationary phase: Silica gel or alumina coated on a glass plate

Flat Methods: Paper

Mobile phase: Liquid solvent

Stationary phase: Cellulose paper

Immunochromatography (Lateral Flow Assay)

Rapid immunochromatographic tests combine chromatographic movement with Ag-Ab specificity.

Sample Pad

Basic Parts of the Cassette

• __________: Where the fluid specimen is dropped; filters and normalizes the sample.

Conjugate Pad

Basic Parts of the Cassette

__________: Contains the labeled antibodies (often gold nanoparticles) that immediately bind the analyte.

Reaction Membrane (Nitrocellulose)

Basic Parts of the Cassette

__________: Contains immobilized antibodies forming the Test and Control lines.

Absorbent Pad

Basic Parts of the Cassette

__________: Located at the end to wick the fluid and maintain capillary action.

Backing Card

Basic Parts of the Cassette

__________: The structural plastic/cardboard holding it all together.

Non-Competitive (Sandwich) Immunoassay

• Pregnancy test (hCG)

• Large antigens with multiple epitopes.

• POSITIVE RESULT: Line appears (analyte bridges labeled Ab and capture Ab).

Competitive Immunoassay

• Drug tests

• Small haptens (drugs, hormones) with one epitope.

• POSITIVE RESULT: Line disappears (analyte blocks labeled Ag from binding).

Heterogeneous Immunoassays

__________:

• Requires physical separation of bound and free components.

• Methods include extensive washing steps and centrifugation.

• Highly sensitive for low-concentration analytes.

Homogeneous Immunoassays

__________:

• Does not require physical separation; the entire reaction occurs continuously in solution.

• Faster testing, easier automation, and no washing steps required.

• Primarily used for therapeutic drugs, hormones, and haptens.

C

Which of the following terms refers to the specific substance being measured in an assay?

• a. Conjugate

• b. Catalyst

• c. Analyte

• d. Isotope

• e. Buffer

B

The initial, thermodynamic force of attraction between a single binding site on an antibody and a single epitope on an antigen is known as:

• a. Avidity

• b. Affinity

• c. Specificity

• d. Sensitivity

• e. Valency

D

What term represents the overall strength of binding between multivalent antigens and multivalent antibodies?

• a. Specificity

• b. Electroosmotic flow

• c. Affinity

• d. Avidity

• e. Pathogenicity

A

In assay methodology, the ability to bind strictly to one specific antigen or antibody without cross-reacting is termed:

• a. Specificity

• b. Avidity

• c. Sensitivity

• d. Affinity

• e. Precision

C

Which term is defined as the lowest limit of detection for an analyte, dealing primarily with identifying very low concentrations?

• a. Affinity

• b. Specificity

• c. Sensitivity

• d. Mobility

• e. Resolution

B

During electrophoresis, which electrode attracts cations (positively charged molecules)?

• a. Anode

• b. Cathode

• c. Diode

• d. Stationary phase

• e. Nitrocellulose membrane

B

The specific pH at which a molecule has a net zero electrical charge and stops migrating during electrophoresis is called the:

• a. Neutrality point

• b. Isoelectric point (pI)

• c. Zwitterion limit

• d. Electrophoretic equilibrium

• e. Buffer pH

C

Which supporting medium used in Serum Protein Electrophoresis (SPE) provides a finer pore structure, better resolution, and is preferred for clinical use?

• a. Cellulose acetate

• b. Silica gel

• c. Agarose gel

• d. Cellulose paper

• e. Fused-silica

E

In Serum Protein Electrophoresis (SPE), which major serum protein typically migrates the fastest and forms the most abundant visible band?

• a. Alpha-1 globulin

• b. Alpha-2 globulin

• c. Beta globulin

• d. Gamma globulin

• e. Albumin

D

A sharp, narrow spike (M-spike) in the gamma or beta region on a Serum Protein Electrophoresis pattern is most indicative of:

• a. Normal protein distribution

• b. Chronic infection

• c. Polyclonal increase

• d. Monoclonal proliferation

• e. Hypogammaglobulinemia

D

Which disease state is characterized by a broad, diffuse increase in the gamma region on an SPE, representing a wide variety of antibodies?

• a. Multiple Myeloma

• b. Waldenström Macroglobulinemia

• c. AL Amyloidosis

• d. Chronic infection or inflammation

• e. Monoclonal Gammopathy of Undetermined Significance

B

What is the primary purpose of performing Immunofixation Electrophoresis (IFE) after an abnormal spike is identified on SPE?

• a. To separate anions and cations simultaneously

• b. To determine the exact type of immunoglobulin causing the spike

• c. To measure the exact concentration of the monoclonal protein

• d. To identify the isoelectric point of the protein

• e. To separate the protein from neutral molecules

C

Which condition is a B-cell lymphoma that produces excess, heavy IgM, leading to hyperviscosity syndrome?

• a. AL Amyloidosis

• b. Multiple Myeloma

• c. Waldenström Macroglobulinemia

• d. Dengue fever

• e. Toxoplasmosis

C

What is a unique advantage of Capillary Zone Electrophoresis (CZE) compared to other methods?

• a. It requires extensive physical washing steps.

• b. It separates neutral molecules efficiently.

• c. It can separate both anions and cations in the same run via electroosmotic flow.

• d. It uses a solid stationary phase for separation.

• e. It detects uncharged large proteins exclusively.

B

In chromatography, what is the term for the component (such as a liquid solvent or inert gas) that carries the sample through the system?

• a. Stationary phase

• b. Mobile phase

• c. Solid support

• d. Absorbent pad

• e. Conjugate pad

D

In a rapid immunochromatographic cassette, which part contains the labeled antibodies (often gold nanoparticles) that immediately bind to the analyte?

• a. Sample pad

• b. Reaction membrane

• c. Absorbent pad

• d. Conjugate pad

• e. Backing card

D

In a lateral flow assay, what is the primary function of the Control Line?

• a. It wicks fluid to maintain capillary action.

• b. It captures the analyte-antibody complexes for a positive result.

• c. It filters and normalizes the initial specimen.

• d. It validates the test by confirming fluid flow.

• e. It provides structural support for the cassette.

C

In a Non-Competitive (Sandwich) Immunoassay, what indicates a positive result?

• a. A line disappears because the analyte blocks labeled Ag from binding.

• b. No line forms at the test region.

• c. A line appears because the analyte bridges the labeled Ab and capture Ab.

• d. The control line fails to appear.

• e. Only the control line appears.

A

In a Competitive Immunoassay, what indicates a positive result (i.e., the presence of the target analyte)?

• a. The test line disappears or fails to form.

• b. The test line appears and becomes darker.

• c. The control line disappears.

• d. Both the test and control lines appear equally strong.

• e. The conjugate pad changes color.

D

Which type of immunoassay does NOT require physical separation of bound and free components, allowing the entire reaction to occur continuously in solution?

• a. Heterogeneous immunoassay

• b. Indirect ELISA

• c. Sandwich immunoassay

• d. Homogeneous immunoassay

• e. Precipitation assay

C

What is the main distinguishing feature of an Enzyme-Linked Immunosorbent Assay (ELISA) compared to broader Enzyme Immunoassays (EIA)?

• a. It requires the use of a radioactive tag.

• b. The reaction occurs continuously in a liquid solution.

• c. It requires a solid phase (e.g., microtiter plate).

• d. It measures the isoelectric point of antigens.

• e. It requires an inert gas mobile phase.

B

What does an Indirect ELISA primarily detect in a patient's serum?

• a. Patient antigens

• b. Patient antibodies

• c. Complement proteins

• d. Phagocytic cells

• e. Exotoxins

A

In infectious disease testing, what does the "C" in the TORCH procedures stand for?

• a. Cytomegalovirus

• b. Coronavirus

• c. Chlamydia

• d. Candida

• e. Campylobacter

C

Which enzyme, acting as a first line of defense in human secretions, functions by directly breaking down bacterial cell walls?

• a. Horseradish peroxidase

• b. Beta-galactosidase

• c. Lysozyme

• d. Prealbumin

• e. Sulfosalicylic acid

E

Which microbial evasion tactic is specifically used by pathogens to prevent phagocytosis by host cells?

• a. Releasing natural killer cells

• b. Triggering the alternative complement pathway

• c. Modifying the host's isoelectric point

• d. Producing IgM

• e. Producing a capsule

D

Which class of immunoglobulin is characteristically increased during helminth (worm) infections?

• a. IgA

• b. IgG

• c. IgM

• d. IgE

• e. IgD

B

Which complement pathway can directly kill parasites by being activated by foreign cell surface components without the need for antibodies?

• a. Classical Pathway

• b. Alternative Pathway

• c. Lectin Pathway

• d. Heterogeneous Pathway

• e. Homogeneous Pathway

A

The mechanism by which effector cells bind to the Fc region of an attached antibody and release lethal cytotoxic granules onto a parasite is called:

• a. Antibody-Dependent Cellular Cytotoxicity (ADCC)

• b. Opsonization

• c. Phagocytosis

• d. Capillary Zone Electrophoresis

• e. Sequestration

E

Which cell is identified as the primary cell responsible for killing parasites, as well as participating in hypersensitivity reactions?

• a. Neutrophil

• b. Basophil

• c. Monocyte

• d. T Cell

• e. Eosinophil

C

What type of immunoglobulin is typically the primary responder produced upon an individual's initial (first) exposure to an infection?

• a. IgG

• b. IgA

• c. IgM

• d. IgE

• e. IgD