Acid Phosphatase

By
Prof. Dr. Azza Mohamed Kamel Abdu Allah
 Acid Phosphatase

• Acid phosphatase (ACP)

belongs to the same group
of phosphatase enzymes
as ALP and is a hydrolase
that catalyzes the same
type of reactions.
• The major difference
between ACP and ALP is
the pH of the reaction.
• ACP functions at an
optimal pH of
approximately 5.0.
 Tissue Source

• ACP activity is found in the prostate, bone, liver, spleen, kidney, erythrocytes, and

platelets.

• The prostate is the richest source, with many times the activity found in other tissue.

Diagnostic Significance

• Historically, ACP measurement has been used as an aid in the detection of prostatic

carcinoma, particularly metastatic carcinoma of the prostate.

• Total ACP determinations are relatively insensitive techniques, as it detects prostatic

carcinoma only when metastasized.

• Newer markers, such as prostate-specific antigen (PSA), are more useful screening and

diagnostic tools
How to isolate prostatic portion of ACP?

• One of the most specific substrates for prostatic ACP is thymolphthalein monophosphate.

• The prostatic fraction of ACP is inhibited by tartrate (Chemical inhibition).

• Serum and substrate are incubated both with and without the addition of l-tartrate.

• ACP activity remaining after inhibition with l-tartrate is subtracted from the total ACP activity

determined without inhibition, and the difference represents the prostatic portion

• Importance 1:-Values are usually normal in the majority of cases and, in fact, may be elevated only in

about 50% of cases of prostatic carcinoma that has metastasized.

• One technique with much improved sensitivity over conventional ACP
assays is the immunologic approach using antibodies that are specific
for the prostatic portion.
• Immunochemical techniques, however, are not of value as screening
tests for prostatic carcinoma.
• PSA is more likely than ACP to be elevated at each stage of prostatic
carcinoma
• PSA is particularly useful to monitor the success of treatment
• PSA is controversial as a screening test for prostatic malignancy
because PSA elevation may occur in conditions other than prostatic
carcinoma, such as benign prostatic hypertrophy and prostatitis.
• Importance 2 :-ACP elevations have been reported in hyperplasia of
the prostate and prostatic surgery.
• There are conflicting reports of elevations following rectal examination
and prostate massage.
• When elevations are found, levels usually return to normal within 24
hours
• Importance 3 :- ACP assays have proved useful in forensic clinical chemistry,

particularly in the investigation of rape.

• Vaginal washings are examined for seminal fluid–ACP activity, which can persist for

up to 4 days.

• Elevated activity is presumptive evidence of rape in such cases.

• Importance 4 :- Serum ACP activity may frequently be elevated in bone disease

associated with the osteoclasts.

• Elevations have been noted in Paget’s disease, in breast cancer with bone

metastases, and in Gaucher’s disease, in which there is an infiltration of bone

marrow and other tissue by Gaucher cells rich in ACP activity.

• Importance 5 :- Because of ACP activity in platelets, elevations are observed when

platelet damage occurs, as in the thrombocytopenia resulting from excessive

platelet destruction from idiopathic thrombocytopenic purpura.

 Assay for Enzyme Activity

• Assay procedures for total ACP use the same techniques as in ALP assays but are performed at an acid
pH:
• p-Nitrophenolphosphate∆ p-Nitrophenol + phosphate ion
• The reaction products are colorless at the acid pH of the reaction, but the addition of alkali stops the
reaction and transforms the products into chromogens, which can be measured
spectrophotometrically.
• Some substrate specificities and chemical inhibitors for prostatic ACP measurements have been
discussed previously.
• Thymolphthalein monophosphate is the substrate of choice for quantitative endpoint reactions.
• For continuous-monitoring methods, α-naphthyl phosphate is preferred.
• Immunochemical techniques for prostatic ACP use several approaches, including RIA,
counterimmunoelectrophoresis, and immunoprecipitation.
• Also, an immunoenzymatic assay (Tandem E) includes incubation with an antibody to prostatic ACP
followed by washing and incubation with p-nitrophenylphosphate.
• The p-nitrophenol formed, measured photometrically, is proportional to the prostatic ACP in the
sample.
 Source of Error

• Serum should be separated from the red cells as soon as the

blood has clotted to prevent leakage of erythrocyte and platelet
ACP.
• Serum activity decreases within 1 to 2hours if the sample is left
at room temperature without the addition of a preservative.
• Decreased activity is a result of a loss of carbon dioxide from the
serum, with a resultant increase in pH.
• If not assayed immediately, serum should be frozen or acidified
to a pH lower than 6.5.
• With acidification, ACP is stable for 2 days at room temperature.
• Hemolysis should be avoided because of contamination from
erythrocyte ACP.
• RIA procedures for measurement of prostatic ACP require non
acidified serum samples.
• Activity is stable for 2 days at 4°C.
Reference Range
• Prostatic ACP, 0 to 3.5 ng/mL
• Tartrate-resistant ACP, adults: 1.5–4.5 U/L (37°C)
children: 3.5–9.0 U/L (37°C)