L3 Clinical Chemistry

🧠 Concepts Introduced

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Plasma Proteins

Plasma proteins are a diverse group of proteins found dissolved or suspended in the liquid portion of the blood (plasma). They make up about 7% to 8% of the total volume of blood plasma.

The three primary categories of plasma proteins are:

• Albumin (~55%–60%): The most abundant plasma protein; it maintains oncotic pressure (fluid balance) and transports fatty acids, steroid hormones, and drugs.

• Globulins (~38%): Divided into alpha, beta, and gamma globulins (antibodies), they handle immune defense and transport iron, lipids, and ions.

• Fibrinogen (~7%): A crucial clotting factor that converts into insoluble fibrin threads to stop bleeding when injuries occur.

Most plasma proteins are synthesized by the liver, with the notable exception of gamma globulins (antibodies), which are produced by plasma B-lymphocytes.

Plasma proteins are distinct from cellular or tissue proteins due to their location, function, and specific specialization:

Location and Environment: Plasma proteins circulate freely inside the blood vessels and extracellular fluid of the circulatory system. Other proteins (often called structural, intracellular, or tissue proteins) reside inside cells (e.g., keratin in hair/skin, actin/myosin in muscle) or are bound to cell membranes.

Primary Functions: Plasma proteins specialize in systemic tasks: regulating blood pH, maintaining blood volume/pressure (osmotic pressure), transporting molecules across the body, fighting infections, and clotting blood. Normal/tissue proteins generally perform localized or structural roles, such as catalyzing metabolic reactions inside a specific cell (enzymes) or building cellular architecture.

Production and Turnover: Most plasma proteins are actively secreted into the bloodstream—primarily by the liver—to serve ongoing, whole-body regulatory roles. Cellular proteins are typically manufactured inside a specific cell for use right there or within that specific tissue

Functions of Plasma Proteins

1. Maintenance of oncotic pressure

proteins capture water to avoid edema

basically cuz of pumping of blood from the heart through vessels (hydrostatic pressure) which pushes fluid and nutrients out of the capillaries into the tissues to nourish cells (water, ions, small nutrients etc.) but not protein cuz proteins are too big

but because the concentration of proteins in the blood vessel is too high which causes water to diffuse into the cell through osmosis (the pulling of fluids back in is called oncotic pressure)

if there is shortage of plasma proteins due to malnutrition, liver disease or kidney loss, blood's oncotic pressure drops significantly so water cannot be captured which causes edema

2. Buffering of pH

3. Transporter

Unlike structural proteins or enzymes that stay in one place, these specialized proteins act as carriers, membrane channels, or receptors to move vital molecules safely through the body.

1. Plasma Carrier Proteins (Soluble Transporters) These proteins travel freely dissolved in the blood plasma to carry molecules that are insoluble or toxic on their own.

• Albumin (Left Panel):
  • What it does: Albumin is the most abundant plasma protein. As shown in the breakdown of hemoglobin, when old red blood cells are destroyed, they form unconjugated bilirubin (which is toxic and insoluble in water).
  • How it works: Albumin acts as a carrier protein, binding tightly to the bilirubin and safely transporting it through the bloodstream to the liver, where it can be processed and excreted.
• Transferrin (Right Panel):
  • What it does: A specialized plasma protein dedicated to carrying iron (Fe^{3+}) through the circulation.
  • How it works: Because free iron is reactive and dangerous to tissues, transferrin binds up to two iron ions safely, acting like a shuttle bus that delivers iron from storage or absorption sites to target tissue cells.

2. Membrane Transporter Proteins (Channels / Exporters) These are embedded directly into the cell membranes to move substances across physical barriers.

• Ferroportin (Right Panel):
  • What it does: Found on the membrane of enterocytes (intestinal cells), it is a transmembrane efflux protein (an exporter).
  • How it works: Ferroportin acts as a gate, pumping iron (Fe^{2+}) out of the intestinal cells and across the membrane into the bloodstream (circulation).

3. Associated Enzymes and Receptors Working hand-in-hand with these transporters are helper proteins that alter the transported substances or help cells catch them:

• Hephaestin & Ceruloplasmin (Right Panel):
  • What they do: These are copper-containing proteins that act as ferroxidases.
  • How they work: As iron exits the cell via ferroportin as Fe^{2+}, these enzymes oxidize it into Fe^{3+}. This chemical conversion is essential because transferrin can only bind to Fe^{3+}.
• Transferrin Receptor / TR (Right Panel):
  • What it does: A receptor protein embedded in the membrane of target tissue cells.
  • How it works: When transferrin carrying iron floats past a tissue cell, the transferrin receptor recognizes it, binds to it, and pulls the iron-transferrin complex into the cell so the tissue can use the iron.

4. Humoral immunity

b-cells. antibody immunity

Most plasma proteins are produced by liver. Immunoglobulins are exceptions Protease on immune response

5. Functional enzymes

Thrombin & factor "?"a are enzymes involved in blood coagulation.

They are not the only functional enzymes. Others include lecithin-cholesterol acyltransferase in esterification of HDL cholesterol & complement C1s/C1r involved in immune defense

Functional Enzymes in the Blood Plasma contains specialized enzymes that circulate to perform specific biochemical tasks, including blood clotting, lipid metabolism, and immune defense.

1. Enzymes in Blood Coagulation (Clotting Cascade)

• The Role of "a": Factors labeled with the letter "a" (such as Factor XIa, Factor IXa, Factor Xa, and Factor Va/VIIIa) represent the active forms of these proteins.
• Proteolytic Enzymes: Active clotting factors and thrombin function as proteolytic enzymes (proteases), acting like molecular scissors to cut and activate subsequent proteins in the cascade.
• Thrombin's Central Role: Thrombin is a key functional enzyme that converts soluble fibrinogen (Factor I) into loose fibrin, forming the mesh network of a blood clot.

2. Other Functional Plasma Enzymes

• Lecithin-Cholesterol Acyltransferase (LCAT): An enzyme involved in lipid metabolism that facilitates the esterification of HDL cholesterol.
• Complement C1s / C1r: Specialized enzymes that are part of the complement system, playing a direct role in innate immune defense by helping identify and destroy pathogens.

Key Distinctions: Enzymes vs. Substrates vs. Cofactors

• Enzymes: The active catalytic proteins that drive reactions (e.g., Thrombin, Factor Xa, LCAT).
• Substrates (Not enzymes): The target molecules that enzymes act upon (e.g., Fibrinogen is the substrate cut by thrombin to form fibrin).
• Cofactors (Not enzymes): Helper proteins that bind to enzymes to speed up reactions rather than performing chemical cuts themselves (e.g., Factor V and Factor VIII).

6. Enzyme inhibitors

AAT & AMG prevents excessive immune-mediated enzymatic damage

Protease on immune response

Enzyme inhibitors (antiproteases) primary job is to act as a safety brake, preventing powerful proteolytic enzymes from causing runaway "friendly fire" damage to healthy tissues during inflammation and immune responses.

1. The Danger of Unchecked Proteases When your immune system fights off an infection, white blood cells (like neutrophils and macrophages) release aggressive proteolytic enzymes—such as neutrophil elastase—to destroy pathogens and clear out cellular debris. However, these enzymes don't discriminate well; they will happily chew up your own healthy structural proteins, such as the elastic fibers in your lungs and blood vessels.

2. How Enzyme Inhibitors Protect Us To prevent this self-destruction, the liver secretes protective antiproteases into the bloodstream:

• alpha_1-antitrypsin (AAT):
  • A major plasma protein belonging to the serpin (serine protease inhibitor) family.
  • It circulates through the blood and neutralizes neutrophil elastase, stopping it from destroying healthy elastic tissues after an immune response is done.
  • (Clinical note: People with a genetic deficiency in AAT cannot stop elastase, leading to chronic lung damage like emphysema, even without smoking).
• alpha_2-Macroglobulin (AMG):
  • Another large plasma protein inhibitor that acts like a molecular "trap" or sponge, physically enveloping and neutralizing a broad spectrum of different proteases circulating in the blood.

3. The Big Picture: Protease vs. Antiprotease Balance As shown on the right side of your slide, the body maintains a strict equilibrium:

• Proteases (like serine, cysteine, aspartic, and metalloproteinases) are constantly produced by cells, platelets, or invading microbes.
• Antiproteases (like Serpins, AAT, antithrombin, AMG, and TIMPs) are the plasma-derived counter-force that neutralizes them to maintain tissue health.

Clinical Indications & Alterations

Indications for protein test

1. Unexplained weight loss
2. Fatigue
3. Edema
4. Liver disease
5. Kidney disease
6. Bone pain

Hyperproteinaemia

Increased protein synthesis (True Hyperproteinaemia): hypergammaglobulinemia, paraproteinaemia

Decreased volume of distribution (Relative Hyperproteinaemia): dehydration, haemoconcentration during venepuncture

Hypoproteinaemia

1. Decreased protein synthesis: malnutrition, malabsorption, liver disease, humoral immunodeficiency ​Malnutrition & Malabsorption: If you don't eat enough protein (malnutrition) or your gut cannot absorb nutrients properly (malabsorption), your liver lacks the raw building blocks (amino acids) needed to synthesize proteins like albumin. ​ Liver Disease: Since the liver is the primary factory that builds most plasma proteins, severe liver damage (like cirrhosis) shuts down protein production. ​ Humoral Immunodeficiency: A condition where the immune system fails to synthesize adequate antibody proteins (immunoglobulins).

2. Increased volume of distribution (dilution): overhydration, increased capillary permeability (septicaemia, hypoxia) ​Overhydration: Excess water intake or fluid retention expands the blood plasma volume, lowering the relative concentration of proteins.

   Increased Capillary Permeability (Septicaemia, Hypoxia): During severe infections (septicaemia) or low oxygen states (hypoxia), the tiny pores in your capillary walls become leaky. Plasma proteins escape out of the blood vessels and into the tissues, dragging water with them and dropping the protein concentration in the blood.

3. Increased loss of protein: nephrotic syndrome, protein- losing enteropathy, burns ​Nephrotic Syndrome: Severe kidney damage causes the microscopic filters (glomeruli) to malfunction, allowing large proteins like albumin to spill out into the urine (proteinuria).

   ​Protein-Losing Enteropathy: Diseases of the gastrointestinal tract that cause proteins to leak directly into the gut.

   ​Burns: Severe skin burns damage blood vessels and cause massive fluid and protein weeping directly through the damaged skin surface.

4. Increased catabolism: sepsis, fever, trauma, malignant disease ​Sepsis, Fever, Trauma, and Malignant Disease: Severe physical stress, widespread inflammation, major injuries, or cancer put the body into a hyper-catabolic state. The body burns through its protein stores at an accelerated rate to fuel the immune response or repair damaged tissues, dragging overall blood protein levels down.

Testing Methods

A. Measurement of total protein

(copper, dye - binding, spectrophotometer)

1. Direct Optical Methods

UV-visible spectrophotometry measures protein concentration using Beer Lambert Law

270-290 nm: Measures tyrosine and tryptophan residues

220-225 nm: Measures peptide bonds; requires removal of interfering small molecules like urea (Because other small molecules (like urea) also absorb light at this low wavelength, they interfere with the reading and must be removed beforehand.)

2. Biuret Method for routine total protein analysis

Reagent: Cupric sulfate (CuSO4) in alkaline solution

Reaction: Cu(II) complexes with lone pair electrons on the nitrogen of amide bonds

Measurement: Absorbance of the blue- colored complex is measured at 540 nm

Interferences: Hemolysis(damaged red blood cells leaking contents into the serum) and lipaemia (high fat/lipid cloudiness in the blood sample)

test results: • Blue (No Change): Means no protein or insufficient peptide bonds are present in the sample. The solution stays the original color of the Biuret reagent (containing copper(II) ions). • Purple / Violet / Mauve: Means proteins or polypeptides are present. Copper(II) ions react with at least two peptide bonds in an alkaline environment to form a colored coordination complex. • Pink / Pinkish-Purple: Can indicate shorter chains like short peptides or digested proteins, or a very low concentration of protein. • Color Intensity: The darkness of the purple color shows the concentration of protein (darker purple means more protein).

B. Measurement of albumin

specific dye - binding This is the standard clinical technique where a specific dye (such as bromocresol green BCG or bromocresol purple BCP) is added to the serum sample. ​How it works: The dye binds specifically to albumin molecules, causing a measurable shift in color intensity that is directly proportional to the albumin concentration.

​More Albumin -> More Dye-Albumin Complexes -> Deeper Color Intensity -> Higher Light Absorbance. ​Less Albumin -> Fewer Complexes -> Fainter Color -> Lower Light Absorbance.

C. Serum protein electrophoresis (SPEP)

major plasma protein pattern

1. Common dyes used to visualise proteins

Coomassie brilliant blue (Bradford method) - stains certain amino acids only like arginine, lysine, histidine, etc.

Ponceau S - Red

Amido black 10B - Black

Silver staining (extremely sensitive) - An extremely sensitive method capable of detecting tiny trace amounts of protein, yielding a black visualization.

2. Buffer for Electrophoretic Separation

Carries the applied current to move the proteins

Sets the pH of the system - influences net charge of the moving proteins

​In an alkaline environment, proteins take on a net-negative charge.
​Because opposites attract, these negatively charged proteins migrate toward the anode (the positive electrode).

​Acidic Environment: In contrast, an acidic environment promotes a net-positive charge, causing proteins to migrate toward the cathode (the negative electrode).

3. Serum Protein Electrophoresis (SPEP) Workflow

1. Place the gel in the electrophoresis chamber.
2. Apply voltage to initiate protein migration.
3. Stain and wash the gel (using dyes like Coomassie, Ponceau S, or Amido Black).

How Staining Works (Step-by-Step) The process of staining a gel usually involves three main stages:

• Fixation (Locking them in place) Right after electrophoresis, the proteins are just loosely sitting in the gel matrix and can easily diffuse or float away. Before adding dye, the gel is treated with a fixative (usually containing an acid and alcohol). This instantly precipitates the proteins, "gluing" them tightly into their specific spots in the gel so they can't move.

• Staining (Binding the dye) The gel is then soaked in a bath containing a protein-binding dye (such as Coomassie Brilliant Blue, Ponceau S, or Amido Black). The chemical interaction: The dye molecules are designed to stick to specific parts of proteins—such as the basic amino acid residues (like lysine, arginine, and histidine) or via hydrophobic interactions. Wherever there is a protein band in the gel, the dye attaches to it and turns it a distinct color (blue, red, or black depending on the dye used).

• Destaining (Cleaning the background) Right after soaking, the entire gel (including the empty spaces where there are no proteins) will look dark and stained. To fix this, the gel is washed in a destaining solution. This washes away all the loose, unattached dye from the empty parts of the gel. It leaves behind a clear, see-through background where only the protein bands retain the dark color, creating sharp, readable lines or peaks.

4. Scan the stained gel using a densitometric scanner.
5. Use density data to produce a viewable tracing curve showing separate peaks.

five distinct peak bands moving from fastest to slowest migration: 
​Albumin: The fastest-moving protein, forming the largest peak closest to the anode because it is relatively small and heavily negatively charged.

​Globulins: Slower-moving proteins that separate into four subsequent fractions:
​Alpha-1 ​Alpha-2 ​Beta ​Gamma (the slowest group, where antibodies sit)

Variables affecting protein electrophoresis

• Analyte charge (mainly)
• Analyte size and shape
• Buffer pH
• Electrical current
• System temperature
• Substrate viscosity

Effects of Buffer pH on Protein Migration

Alkaline Buffer: Most common for serum protein electrophoresis applications. Alkaline Environment: Promotes a net-negative charge on proteins, causing them to move toward the anode (positive electrode).

Acidic Environment: Promotes a net-positive charge on proteins, causing them to move toward the cathode (negative electrode).

Resulting Fractions (Plasma Protein Pattern): Proteins separate into five distinct peak bands from fastest to slowest migration:

• Albumin (The largest peak closest to the anode)
• Globulins (Split into alpha 1, alpha 2, beta, and gamma fractions)

D. Specialised Immunology & Particle Assays

Highly sensitive tests that use antibodies to find specific targets instead of general protein structures.

1. Immunofixation

detect specific immunoglobulins & light chains

to identify and characterize specific monoclonal (artificial clone) immunoglobulins (antibodies) and light chains in a patient sample.

​Step-by-Step Immunofixation Workflow
​Sample Application: Multiple aliquots of the exact same patient sample are applied side-by-side onto an agarose gel. 

​Protein Separation: An electric current is applied, causing the serum proteins to separate by electrophoresis into distinct tracks/lanes across the gel. 

​Antibody Overlay: Instead of standard total protein staining, specific antibody solutions (antisera) are soaked into strips and overlaid onto each individual lane. These antibodies target specific heavy chains (mu, gamma, alpha) and light chains (lambda, kappa). 

​Precipitation & Detection: If the matching target protein is present in that lane, the added antibody binds to it, forming an insoluble immune complex that precipitates into a visible band. The gel is then washed, stained, and analyzed.

2. Immunonephelometry & immunoturbidimetry

Immunoturbidimetry: Detector at 180 degrees to incident light. Best for quantifying larger particles

Immunonephelometry: Detector at an angle (usually 90 or 45 degrees) to incident light. Best for quantifying smaller particles

two techniques to measure the exact concentration of specific proteins (like immunoglobulins, complement factors, or albumin) in a fluid sample. ​Both methods rely on an antigen-antibody reaction: when you mix a patient's sample containing a target protein (antigen) with specific antibodies, they bind together to form microscopic immune complexes. These complexes make the liquid cloudy and cause it to scatter light. ​The main difference between them lies in how and where the detector measures that light.

Limit of detection: 10 mg/L or lower

cuvette (small, clear, specialized glass, quartz, or plastic tube/container used to hold liquid samples)

Immunoturbidimetry ​Detector Position: Placed directly in a straight line at 180 degrees to the incident light source (measuring what passes straight through the sample). 

​Best For: Quantifying larger particles / aggregates. 

​How it works: As light passes through the cuvette, large protein clumps block and scatter the light beam. The detector measures the reduction in transmitted light. The cloudier the sample, the less light reaches the detector. ​Because the cloudiness intercepts the light, fewer photons make it straight through to the other side. ​The detector measures this drop—calculating how much light failed to pass straight through because the protein clumps blocked or scattered it.

Immunonephelometry ​Detector Position: Placed at an angle (usually 90 degrees or 45 degrees) to the incident light beam. 

​Best For: Quantifying smaller particles. 

​How it works: Instead of looking at the light that passes straight through, a nephelometer measures the light that is scattered sideways or forward-angled by the immune complexes. 

​Why it's unique: Measuring scattered light against a dark background provides a very high signal-to-noise ratio, making it exceptionally sensitive for low-abundance proteins.

3. ELISA (Immunoassay) L4 Clinical Chemistry#Mass Assays (Immunoassays)

for extremely low concentration analyte via highly targeted antibody binding and enzyme amplification steps.

highly targeted antibody binding like immunifixation and enzyme amplification

​The most common type of ELISA is the "Sandwich ELISA," which works like building a molecular sandwich:

​The Capture: The bottom of a plastic well (in a microplate) is coated with antibodies designed to grab and hold the target protein.

​The Sample: The patient's fluid sample is added. If the target protein is present, the capture antibodies catch it and hold it in place. (Everything else is washed away).

​The Detection Antibody: A second antibody—linked to the amplification enzyme—is added. It binds to the top of the trapped protein, completing the "sandwich."

​The Substrate (The Readout): A colorless substrate liquid is poured in. The enzyme instantly reacts with it, changing the liquid's color (e.g., turning bright yellow or blue).

​The Measurement: A spectrophotometer measures the intensity of the color. The darker the color, the more target protein was in the original sample.

Total protein in plasma

60-80 g/L in serum

100 proteins that have a physiological function in the plasma

Two most abundant proteins: Albumin & immunoglobulin

Albumin:

• Synthesized in liver
• MW 66kDa
• 35-55 g/L
• 60%-70% of total serum proteins
• Decreased during acute phase reaction

A/G Ratio

Globulin = Total protein - Albumin

Albumin / Globulin ratio (ref interval: 1 - 2)

Low A/G: Overproduction of globulin in autoimmune & multiple myeloma; selective loss of protein in nephrotic syndrome; lower production of albumin in liver cirrhosis

High A/G: Lower production of globulin in leukemia, genetic deficiencies in globulin production; underuse of albumin in hypothyroidism

Serum Protein

Serum protein electrophoresis Limit of detection: 1 g/L

Urine Protein

Higher MW proteins normally do not pass through glomerular filter

Renal tubular reabsorption is inversely related to molecular size

Smaller proteins like albumin & light chain are reabsorbed in renal proximal tubule

Fractional Analysis: The Five SPEP Zones

Albumin

Synthesized in liver MW 66kDa 35-55 g/L 60%-70% of total serum proteins Decreased during acute phase reaction

Function:

• Maintains plasma oncotic pressure
• Buffer for acid-base balance
• Transports ligands like bilirubin, free fatty acids, hormones (thyroxine, cortisol), Ca, Mg, drugs (warfarin, penicillin)
• Endogenous source of amino acids

Clinical conditions

Hyperalbuminemia: Little clinical significance Dehydration Artefactual: infusion of plasma or albumin- containing solution

Hypoalbuminemia: Increased catabolism: tissue damage & inflammation - sepsis, fever, trauma, malignant disease Decreased synthesis: liver disease, decreased protein intake, malabsorption Increased loss of protein: nephrotic syndrome, protein-losing enteropathy, burns

Alpha - 1 Globulins

Alpha1-antitrypsin (AAT) (1 -3 g/L):

• Major alpha1 protein
• Positive APR, increase in pregnancy & oral contraceptive use
• Antiprotease activity, resulting in neutralizing leukocyte elastase and collagenase
• Deficiency results in lung emphysema and liver disease

#6. Enzyme inhibitors

Alpha - 2 Globulins

Alpha2-macroglobulin (AMG) (1.0 - 2.8 g/L):

• Largest plasma protein

• Protease inhibitors

• Inhibitor of coagulation & fibrinolysis

• Levels up 10x in nephrotic syndrome

• To compensate for the protein loss, liver increases protein synthesis

• Levels increase in acute phase response

Beta Globulins

LDL (< 1 g/L; can rise to 4.9 g/L in diseased condition)

Transferrin (TRF) (2.4 - 4.5 g/L):

• Major component of beta-globulin

• Synthesized in liver

• Iron transport: two iron binding site, in ferric form

• Elevated in iron-deficiency anemia

• Negative APR

• Lowered in iron-overload, protein-losing syndrome, severe liver disease, inflammation

• Atransferrinemia: Rare genetic condition. Iron overload in heart and liver. Heart failure and liver cirrhosis

#3. Transporter

• When Transferrin is Too High (e.g., Iron-Deficiency Anemia):
  • ​Why it's bad: While high transferrin is the body's attempt to scavenge every drop of remaining iron, it means there is an absolute shortage of iron available. Without enough iron, the bone marrow cannot manufacture hemoglobin inside developing red blood cells, resulting in microcytic hypochromic anemia (meaning the red blood cells are too small and pale, leading to chronic fatigue, weakness, and poor oxygen delivery throughout the body).
• ​When Transferrin is Too Low (e.g., Liver Failure, Nephrotic Syndrome, or Inflammation):
  • ​Why it's bad: If transferrin levels drop drastically, the blood loses its capacity to safely transport iron. This can lead to un-bound or poorly bound iron circulating dangerously, or it contributes to anemia of chronic disease because iron cannot be efficiently delivered to where it is needed. Furthermore, because transferrin is a negative acute-phase reactant and a major plasma protein, low transferrin often signals severe underlying inflammation, malnutrition, or failing liver function.

Gamma Globulins

Immunoglobulin Synthesized by plasma cell Often can be found throughout the electrophoretic spectrum

Immunoglobulin G (IgG):

• Most abundant, 70-75%
• Only IgG crosses placenta
• 7-16 g/L
• Monomer
• Elevated in liver disease, multiple myeloma, parasitic diseases, and rheumatic diseases

Immunoglobulin A (IgA):

• 10 - 15%, 2nd most abundant
• Dimer
• 0.6 to 3.0 g/L
• SPE: Anodal to IgG
• Secretory IgA is found in tears, sweat, saliva, milk, colostrum, and gastrointestinal (GI) and bronchial secretion
• Role of IgA in plasma is unclear

Immunoglobulin M (IgM): 5 - 10% Ig, 3rd most abundant 0.4-2.3 g/L Pentamer Cannot cross placenta Produced at earlier stage of B-cell development Acute exposure to immunogen or pathogen Monoclonal elevation in Waldenstrom's macroglobulinemia

Clinical Relevance of Ig

if polyclonal the peak is rounded

liver IgA

Clinical Electro-Gammopathy & Disease Profiles

Clinical Indications for in Multiple Myeloma Screening

Suspected multiple myeloma (malignant plasma cells in the bone marrow)

Waldenström macroglobulinemia (malignant B-cell lymphoplasmacytic lymphoma producing monoclonal IgM)

primary amyloidosis (abnormal protein folding and tissue deposition), or related disorder

Unexplained peripheral neuropathy

New-onset anemia associated with renal failure/insufficiency and bone Tesion (hypercalcemia & unexplained bone fracture on radiograph) ​C (Calcium elevation): Bone destruction releases calcium into the blood.
​R (Renal failure/insufficiency): Impaired kidney function driven by light chain toxicity or hypercalcemia.
​A (Anemia): Malignant plasma cells crowding out the bone marrow, suppressing normal red blood cell production.
​B (Bone lesions / lytic lesions): when a patient presents with unexplained anemia alongside failing kidneys and bone issues, screening for myeloma is mandatory.

Back pain in which multiple myeloma is suspected

Rouleaux formation noted on peripheral blood smear

Renal insufficiency with associated serum protein elevation

Bence Jones proteinuria - about 60% cases

Classic Electrophoretic Trajectories (M- Spikes and Bridges)

check the tube colour

Summary

• Proteins in the blood plasma have multiple importantphysiological role

• Plasma a proteins are abundant in albumin and globulins

• Globulins can be further classified into alpha, beta and gamma globulins

• The 'abnormal' levels of each plasma fraction on the serum protein electrophoresis is a useful indicator for the nature of a disease