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Chemical Pathology > Plasma Proteins/Electrophoresis > Flashcards

Plasma Proteins/Electrophoresis Flashcards

1
Q

Immunoglobulins are made from?

A

B-lymphocytes

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2
Q

Complement system are made by?

A

Macrophages

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3
Q

Plasma proteins are made from?

A

Hepatocytes

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4
Q

Measurement of proteins include the measurement of what parameters ?

A
  • Total protein
  • Plasma Albumin
  • C-reative protein
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5
Q

The amount of proteins in the vascular compartment depends on?

A
  • The balance between the rate of synthesis and catabolism or loss
  • Relative distribution between the intravascular and extravascular compartments
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6
Q

Concentration of plasma depends on ?

A

The relative amounts of proteins and water

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7
Q

Plasma proteins are taken up by what process and into what cells during their catabolism ?

A

They are taken up by Pinocytosis into the capillary endothelial cells where they are catabolized

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8
Q

Small chain molecular weight proteins are lost passively though what parts of the body?

A
  • The renal glomeruli
  • The intestinal wall
    **some are reabsorbed with by the Renal tubular cells or after digestion in the intestinal lumen
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9
Q

Functions of plasma proteins?

A

1) Peptide hormones and blood clotting factors
2 ) control of extracellular fluid distribution
3) Trabdport
4) inflammatory response and control of infections

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10
Q

How does plasma protein function as a transporter?

A

Albumin and specific binding proteins transport hormones, vitamins, lipids, bilirubin, calcium, trace metals and drugs

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11
Q

How does plasma protein function in inflammatory response and control of infection?

A

Immunoglobulins and the complement proteins together with CRF and other APRs form part of the immune system and each are involved with inflammatory response

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12
Q

Methods of assessing plasma proteins?

A
  • Plasma proteins may be expressed as either concentration (g/L)
  • Plasma proteins may be assessed based on their functions (prothrombin for clotting, or enzyme function )

1) Measurement of Total protein concentration
2) Assay of specific proteins (based on function
3) Quantify serum proteins by serum electrophoresis

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13
Q

In what case does an abnormal form of protein become present at normal concentration ?

A
  • C1 inhibitor of the classical pathway
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14
Q

What can cause raised Total plasma protein?

A
  • Loss of protein free fluid
  • Excessive stasis during venepuncture
  • Major increase in the constituent of one or more of the immunoglobulins (paraproteins to)
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15
Q

What can cause low total plasma proteins ?

A
  • Dilution
  • hypoalbuminemia
  • immunoglobulin deficiency
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16
Q

Qualitative methods for studying plasma and urinary proteins include the use of what technique ?

A

Electrophoresis

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17
Q

What is electrophoresis? ATIBA

A

Electrophoresis is a technique that separates compounds such as proteins into various components according to their electrical charge

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18
Q

Steps in electrophoresis

A
  • Application of a small amount of serum to a strip of cellulose acetate or agarose gel
  • Passage of a current across the supporting medium for a standard time
  • serum protein is then separated into a1, a2, beta, gamma globulins, and albumin groups
  • Each of the globulins contains several proteins.
  • distinguished after protein staining with dye
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19
Q

The most obvious band in electrophoresis includes ?

A

Albumin - A single protein, makes up the most obvious band

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20
Q

What proteins does alpha 1 globulin band contains ?

A
  • Alpha 1 anti-trypsin
  • Alpha 1 anti-chymotrypsin
  • Alpha 1 acid-glycoprotein
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21
Q

Alpha 2 globulin consists of what proteins ?

A
  • Alpha 2 - macroglobulin
  • *Haptoglobin
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22
Q

Beta globulin contains what proteins ? (B1 & B2)

A
  • Beta 1: Transferrin, LDL
  • Beta 2: C3 complement protein
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23
Q

Gamma globulins contain what proteins ?

A

They are immunoglobulins

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24
Q

Apart from gamma globulin, immunoglobulins can also be found in what other band?

A

Beta and alpha 2*

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25
Q

What happens if you use plasma for electrophoresis instead of serum?

A
  • Fibrinogen appears as a distinct band in the Beta-gamma region
  • May make interpretations difficult
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26
Q

What happens to alpha 2 band In vivo hemolysis and in vitro hemolysis?

A

Alpha 2 band contains Haptoglobin
- In ViVo hemolysis
Alpha 2 band is reduced**
- in Vitro hemolysis
**Alpha 2 band may split into two*

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27
Q

Electrophoretic pattern of acute phase reaction ? (Acute inflammatory reaction)

A
  • There’s stimulation of synthesis of Acute phase proteins
    A rise in alpha 1 globulin and alpha 2 globulin fractions
  • there’s a rise in ESR and plasma viscosity
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28
Q

Electrophoretic pattern in chronic inflammation?

A
  • An increase in immunoglobulin synthesis hence; (diffuse rise)
    Gamma globulin band is INCREASED
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29
Q

Electrophoretic pattern in Liver cirrhosis ?

A
  • Albumin band is REDUCED
  • Alpha 1 band is REDUCED
  • gamma globulin is markedly INCREASED
    Increased plasma IgA concentrations
  • FUSION of beta and gamma bands
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30
Q

Electrophoretic pattern in nephrotic syndrome

A

Depends on severity of the renal lesion
- Early cases:
low plasma albumin concentration may be the only finding
- Established cases;
Albumin band is REDUCED
Alpha 1 band is REDUCED
Gamma globulin band is REDUCED
- increase in HMW alpha 2 macroglobulin;
Alpha 2 band is INCREASED

31
Q

Electrophoretic patter
In Nephrotic syndrome (SLE )?

A

Gamma globulin band may be increased for normal

32
Q

Electrophoretic pattern In alpha 1 anti-trypsin deficiency

A

* Absence or obvious reduction in alpha 1 band*
- A1 antitrypsin variants;
*split alpha 1’ band

33
Q

Normal biological Half life and weight of albumin?

A

HL; About 21 days
Weight : 65Kda

34
Q

Inherited abnormalities of albumin synthesis ?

A

1) Bisalbuminaemias
2) Analbuminaemis

35
Q

Busalbuminaemia ?

A

Two forms of albumin are present

36
Q

Analbuminaemia?

A

Deficient synthesis of protein

37
Q

Causes of hypoalbuminaemia (5)

A

1)Dilutional hypoalbuminaemia
2)Redistribution of albumin
3) decreased synthesis of albumin
4) increased loss of albumin from the body
5) increased catabolism of albumin

38
Q

Dilutional hypoalbuminaemia cases ?

A
  • Result from artefactual changes - Blood sample taken from arm of IV infusion
  • Administration of an excess of protein free fluid
  • Fluid retention in edematous states or during late pregnancy
39
Q

Redistribution of albumin from plasma into the interstitial fluid space is caused by?

A
  • Recumbency
    Plasma albumin concentrations may be 5-10g lower in the recumbent position than In the upright position
  • increased capillary membrane permeability
    Found in most illnesses, in inflammation also or post operatively
40
Q

Decreased synthesis of albumin may be caused by?

A
  • Liver cirrhosis; impairment of synthesis
  • Malabsorption; impaired absorption of dietary peptides and amino acids
  • undernutrition; inadequate supply of Dietary nitrogen
41
Q

How many percent of the body’s albumin is replaced each day?

A

4 percent

42
Q

I’m creased loss of albumin from the body can be caused by?

A
  • In increased membrane permeability
  • The skin; extensive burns or skin disease (psoriasis)
  • **Intestinal wall; protein losing enteropathy
  • The glomeruli; nephrotic syndrome
43
Q

A large part of interstitial fluid is subcutaneous in ?

A

Increased capillary permeability

44
Q

Increased catabolism of albumin can be caused by?

A

Sepsis
Hyperthyroidism
Associated with nitrogen loss

45
Q

Consequences of Hypoalbuminemia

A

Fluid distribution; Edema
Binding functions; Albumin binds calcium, bilirubin and free fatty acids. Hence there’s a poor transport of albumin binding hormones

46
Q

M proteins are also called ?

A

Paraproteins

47
Q

What are paraproteins?

A

Appearance of abnormal, narrow dense band on the electrophoretic strip.
Can be shown to be monoclonal

48
Q

Paraproteins are most commonly found where ?

A

Gamma region of the electrophoretic strip

49
Q

Causes of paraproteinemia ?

A
  • Myelomatosis (most cases)
  • Waldenstrom’s macroglobulinaemia
  • B cell lymphomas (CLL)
  • Cryoglobulinaemia
  • SLE, chronic inflammation, infection
50
Q

Consequences of paraproteins ?

A
  • Increased plasma viscousity
  • Pseudohyponatremia (space occupying effect of the protein)
  • retinal vein thrombosis
  • cerebral thrombosis
  • peripheral gangrene
51
Q

Globin concentration is defined as?

A

The differences between the total plasma protein concentration and albumin concentration

52
Q

What are bence jones proteins

A

Consists of free monoclonal Light chains, found in the urine of patients with B cell malignancy

53
Q

Causes of hypogammaglobulinaemia

A

1)Bruton’s or swiss-type agammaglobulinaemia
2) diabetes mellitus
3) Cushing’s syndrome
4) CKD
5) steroids

54
Q

Consequences of immunoglobulin deficiency?

A

IgA - recurrent respiratory tract infections
IgG - pyogenic infections in lungs and skin (tissue spaces) from toxin producing organisms (staph and strep)
IgM- septicemia

55
Q

In chrons disease what Ig is increased?

A

IgA - it is produced submucosallyv

56
Q

Define electrophoresis - IDOWU

A

Electrophoresis is the migration of charged solutes or particles in a liquid medium under the influence of an electric field

57
Q

Iontophoresis is defined as ?

A

The migration of small ions

58
Q

Zone electrophoresis is defined as?

A

The migration of charged macromolecules in a porous support medium such as paper, cellulose acetate or agarose gel film

59
Q

The positive electrode is the?

A

Anode

60
Q

The negative electrode is the ?

A

Cathode

61
Q

Ampholyes (zwitterion) are positive in what solutions?

A

In solutions more acidic than it’s iso-electric point, and migrates towards the cathode

62
Q

The result of the zone electrophoresis is called?

A

Electrophoretogram - consists of sharply separated zones of a macromolecule

63
Q

The rate of electrolyte mobility is determined by?

A

1) The net charge of the molecule
2) Size and shape of the molecule
3) Strength of the electric field
4) properties of the supporting medium
5) Temperature of operation
6) Viscosity of the buffer solution

64
Q

Components of an electrophoresis machine ? Mention 9

A
  • Electrophoretic tank
  • The electrodes (anode and cathode)
  • The supporting medium
  • The buffers
  • Power supply
  • The specimen applicator
  • Stains
  • Densitometer
  • Comuputer (automated systems)
65
Q

Types of electrophoresis ?

A
  • Zone electrophoresis
  • Isotachophoresis
  • Isoelectric focusing
  • Two dimensional electrophoresis
  • Capillary electrophoresis
66
Q

Subtypes of zone electrophoresis

A
  • Agarose gel electrophoresis
  • Cellulose acetate electrophoresis
  • polyacrilamide gel electrophoresis
67
Q

Dyes used in electrophoresis ? Mention 6

A
  • Ponceau S dye
  • Amido black
  • Oil red O
  • Fat red
  • Sudan black
  • CBB
68
Q

stepwise procedure in electrophoresis ?

A
  • Sample is soaked in a hydrated medium for about 5 mins
  • supporting medium is put into the electrophoresis chamber (that was previously filled with the buffer)
  • A constant voltage is applied to the supporting medium for a specific time
  • support is then removed and placed in a fixative/rapidly dried to prevent diffusion of the sample
  • this is followed by staining the zones with appropriate dye.
69
Q

What’s the function of the buffer solution in electrophoresis ?

A

It must be added to the chamber to maintain contact with the supporting medium

70
Q

Purpose of putting the supporting medium in a fixative before staining?

A

To prevent diffusion of the sample

71
Q

Factors influencing the uptake of dye of the sample provided ?

A

The uptake is dye of the sample is proportional to the sample concentration

72
Q

Simplest way to accomplish detection in electrophoresis is ?

A

Visualization under UV light

73
Q

The most reliable and common way for quantitation in electrophoresis is through ?

A

Densitometry

74
Q

Clinical applications of electrophoresis ?

A
  • Used to establish several clinical conditions
  • used to classify lipoproteins: chylomicrons - origin
    VLDLP - pre Beta band
    IDL - between beta and pre beta bands
    LDL - Beta band
    HDL - Alpha band
  • it is a component of other techniques like southern, northern (RNA) and western (protein) blotting

Chemical Pathology (8 decks)

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