CA2119096A1 - Lyophilized control or reference plasma or control or reference serum for diagnostic purposes - Google Patents
Lyophilized control or reference plasma or control or reference serum for diagnostic purposesInfo
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- CA2119096A1 CA2119096A1 CA 2119096 CA2119096A CA2119096A1 CA 2119096 A1 CA2119096 A1 CA 2119096A1 CA 2119096 CA2119096 CA 2119096 CA 2119096 A CA2119096 A CA 2119096A CA 2119096 A1 CA2119096 A1 CA 2119096A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
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- Sampling And Sample Adjustment (AREA)
Abstract
ABSTRACT OF THE DISCLOSURE:
There is disclosed a lyophilized control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular apo-B-containing lipoproteins, which has a water content of between 1 and 10 %, preferably between 1.5 and 5 %, and is reconstitutable to a clear solution exhibiting a maximum turbidity of 70 LSU. Furthermore, there are disclosed a method of preparing a lyophilized control or reference plasma or control or reference serum as well as a method of preparing a control or reference plasma or control or reference serum.
There is disclosed a lyophilized control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular apo-B-containing lipoproteins, which has a water content of between 1 and 10 %, preferably between 1.5 and 5 %, and is reconstitutable to a clear solution exhibiting a maximum turbidity of 70 LSU. Furthermore, there are disclosed a method of preparing a lyophilized control or reference plasma or control or reference serum as well as a method of preparing a control or reference plasma or control or reference serum.
Description
This invention relates to a lyophilized control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular apo-B-containing lipoproteins, as w~ll as a method of preparing the same.
The lyophilization of aqueous solutions is a common method of producing preparations that are stable during long-term storage. In doing so, the water is removed from the frozen solution by sublimation under vacuum. The water content of storage-stable lyophilisates, in general, amounts to less than 1 %
(w/w). Yet, rendering aqueous solutions storabLe by lyophilization is suitable only if the physico-chemical properties of the ingredients will not be substantially changed by the removal of water~
The lyophilizatlon of lipoprotein-containing plasma or serum constitutes a great problem, since, according to the experience, turbid solutions are obtained upon reconstitution. Lipoproteins may be denatured by lyophilization such that their determlnation by means of electrophoretic or immunologic or optical methods is difficult. A common ; ~ -measure for the turbidity of a solution is LSU (light scattering unit). In general, a solution is considered turbid if it has a measured signal of more than 70 LSU
when measured in the nephelometer at 340 nm and a path length of 1 cm (reference: water).
From the document Clin. Chem. (1990), 36, 366-369, ~. ,, . ' .
`:~
it is apparent that lyophilized serum is not suitable as a reference for determining lipoproteins because of the denaturation of lipoproteins. This holds, in particular, for apo-B lipoproteins, whos~ structure is ., .~
changed during dehydration and rehydration in such a manner that their solubility is reduced, which results in the turbidity of the serum.
Attempts to prevent the formation of turbidity by the addition of stabilizers, such as, e.g., sucrose, to the starting material (serum) have been described in AT B-361,135 and in US-A-4,701,417. Accordingly,, the turbidity, expressed by the LSU measured in - ~
nephelometry, can be inadmissikly high such that these -preparations cannot be applied as references for nephelometric determination. A solution is considered suitable for use in diagnostic methods i it has a maximum turbidity of 70 LSU.
A special method of lyophilization is described in US-A-3,928,566 and US-A-3,932,943. Frozen droplets are formed in the serum or plasma by a sprayin~ procedure, thereby causing an increase in turbidity upon reconstitution of less than 20 ~. However, this method involves high expenditures.
It is the object of the invention to avoid the drawbacks of lipoprotein-containing lyophilisates ~ ~
mentioned above and to provide a lyophilized co~trol or ~ ~ `
reference plasma or control or reference serum for `~
diagnostic purposes, which is capable o~ being reconstitued to a clear solution and can be produced by a simple method.
In accordance with the invention, this object is achieved b~ a lyophilized control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular apo-B-containing lipoproteins, which is characterized in tha$ it has a water content ranging between 1 and 10 ~, preferably between 1.5 and 5 ~, and is reconstitutable to a clear solution exhibiting a maximum turbidity of 70 LSU.
A particularly preferred range of the water content of the lyophilisate is between 2 and 5 %.
It has been shown that, due to a given residual moistura in the lyophilisate, reconstitution to a clear solution is possible. The water content in the lyophilisate is indirectly related to the increase in turbidity by lyophilization. The higher the water content of the hydrated lyophilisate, the low~r the increase in turbidity. A residual moisture of between 1 and 10 ~ (w/w) is suitable primarily because the storage stability may be jeopardized at a higher water content. ~;~
It has also been found that the increase in -~
turbidity due to lyophilization and reconstitution is ~ -r ev~n larger if already the starting material is relatively turbid. Hence, it must be taken care that it is depar~ed from a suitable starting material in order to measu~e not more than 70 LSU in the reconstituted -::
solution. Values of above 70 LSU correspond to a turbidity that renders the solution practically unsuitable for use as a reference or control in nephelometric determinations. Values of up to 50 LSU
are particularly suitable.
The lyophilized control or reference plasma, or ; -control or reference serum according to the invention, ;~
furthermore, may contain non-reducing sugars, such as sucrose, for stabilization. A content of sucrose that ~ `
does not exceed 25 ~, preferably ranging between 12 and 20 %, is advantageous. ;~
The plasma or serum according to the invention is sufficient'y stable for long-term storage. This was not to be foreseen, because the water content was generally reduced in a lyophilisate as far as possible in order ; ;~
to improve the s~orage stability.
The invention, furthermore, relates to a method of preparing a lyophilized control or reference plasma or control or reference serum which is characterized in that lipoprotein-containing plasma or serum optionally is admixed with a non-reducing sugar, preferably sucrose, and is lyophilized until reaching a water content of between 1 and 10 ~
It has been shown that a turbidity of more than 70 LSU in the reconstituted solution can be prevented if -the lyophilisate is incubated in the moist sta~e or in a moist atmosphere. In doing so, the lyophilisate reabsorbs moisture in a reversible manner, which, in turn, results in a clear reconstituted solution. Thus, lyophilisa~es that would yield turbid and hence unusable solutions upon con~entional reconstitution (addition of water) are capable of being reconstituted to solutions ha~ing a turbidity of less than 70 LSU iand thus being usable in diagnostics.
A preferred method of preparing a lyophilized control or reference plasma or control or reference serum, therefore, is characterized in that a lyophilized lipoprotein-containing plasma or serum is incubated in a moist atmosphere. ~ ~;
Another embodiment of the method of preparing a lyophilized control or reference plasma or control or reference serum consists in that a lipoprotein-containing plasma or serum optionally is admixed with non-reducing sugars, preferably sucrose, and is lyophilized and the lyophilisate is adjusted to a water ontent of between 1 and 10 ~ by incubation in a moist atmosphere.
; Thus, con~rol or reference plasmas or control or reference sera that exhibit a maximum turbidity of 70 LSU even can be prepared from lyophilisates having low watericontents, by incubation in a moist atmosphere.
~`~ A method of preparing a control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular, apo-B~
containing lipoproteins is characterized in that a -lyophilized control or reference plasma or control or - 5 - ;
y i ~ :
reference serum according to the present invention is : :
reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU.
A particularly preferred embodiment of the method of preparil~g a control or reference plasma or control or reference serum for diagnostic purposes containing ~ ~
lipoproteins, in particular, apo-B-containing ~ :
lipoproteins consists in that a lyophilized lipoprotein-containin~ plasma or serum is adjusted to a water content of between 1 and 10 ~ by incubation in a moist atmosphere and subsequently is reconstituted to a clear solution exhibiting a maximum turbidity of 70 ~ -.
LSU. ~ ;
Incubation in a moist atmosphere is effected at appropriate temperatures ranging from 4 to 50C, preferably from 30 to 40C, in particular, at 37C, ~: ;
until the turbidity of the reconstituted solution;~
corresponds to a maximum value of 70 LSU. The duration of Lncubation is a funation of the temperature and may last from 1 to 50 da~s, pre~erably from 7 to 28 days.
: The addition of non-reducin~ sugars, in particular : suorose, may be effected prior to lyophilization~
' The invention wlll be explained in more detail by :~ the following examples.
ExampIe 1 Preparation of plasma lyophilisates having different water contents --~
: Rethrombinized fresh plasma samples were mixed : ;: .''.', ~,`
; - 6 -~ .
with 15 % sucrose and examined for turbidity. After this, samples with 20 LSU were lyophilized to a residual moisture of 0.5, 1.1, 3.8 and 8.5 % (w/w) water. For comparison, plasma samples with 48 LSU were lyophilized to a residual moisture of l.1 and 2.8 ~
(w/w). The water content was measured according to Karl Fischer's technique.
The lyophilisates were reconstituted with water and diluted until reaching their starting volumes. The turbidities of the reconstituted solutions were measured by an IMMUN0 video nephPlometer (IMMUN0 AG).
Table 1 indicates that the turbidity of the solution of the lyophilisate having a water content of 0.5 % (w/w) is inadmissibly high. Likewise, it is ;
apparent that plasma exhibiting an elevated self~
turbidity brings about a still admissible turbidity in ~-the reconstituted solution only at an elevated water content of the lyophilisate. There is a linear relationship between the logarithm of the water content and the logarithm of the ratio of the turbidity exhibited by the reconstituted solution to that exhibited by the starting plasma (r = -0.833.
; ' ;'' . '' '~ - ' Table 1 Influence of water content in plasma lyophilisate on turbidity in reconstituted solution Water content inTurbidity in Turbidity in lyophilisatestarting plasmareconstituted (~, w/w) (LSU) solution ~LSU) 0.5 20 100 1.1 20 70 ~:
3.8 ZO ~6 8.5 20 19 1.1 48 ~00 2.8 48 69 Example 2 ~;
Assessment of the storage stability of a control serum as a function of its residual moisture Lp(a)-containing control sera prepared accordi~g to the method of AT-B-361,135 and having a content of sucrose (15 ~) and differeht contents of water were ncubated at 37C for 14, 21 and 28 days. According to : experience, 28 days at 37C correspond to a storage , stability of 2 years at +4C. To assess its stability, Lp(a) (lipoprotein a) was quantitatively determined and the quotient QLp(a) was cal~ulated.
" ,, : Lp(a)-contentn days 37C
QLP ~ a) =-~ ~~
Lp(a)-co~entO days 37C ~ ;
.~
Table 2 Storage stability of control serum having varying water contents ' Water content QLP( a) after an incubation time of (%) 0 d 14 d 21 d 28 d .
3.0 1.00 1.00 1~00 1~04 : ~ ~ :
1.5 1.00 0.94 1.06 l~00 1.0 l.00 1,07 1.05 1.04 0.5 l.00 0.94 0.91 0.94 For the period of time investigated, the storage - `~
stability of the Lp(a)-containing serum is independent of its residual moisture. :
Example 3 Preparation of serum lyophilisates having varying sucrose contents Lp(a)-containing control serum prepared according to the method of AT-B-361,135 was mixed with varying portions of sucrose (S %, 10 %, 15 %), frozen and lyophilized in a manner that the water content varied in the end product. -, ~.
;,~
~:
Table 3 Influence of sucrose content in serum lyophilisate ~ :
on turbidity of reconstituted solution Turbidity (LSU) in reconstituted solution of serum lyophilisate having a sucrose content of Water content (%) 5 % 10 % 15 % ; . `~
sucrose 8.0 - 8.4 2~0 LSU 57 LSU n.d.*
2,9 - 4,2 195 LSU 95 LSU 41 LSU
1,0 - 1.2 _ 210 LSU 194 LSU , 73 LSU ~
*) .... not determined ~ .
Example 4 - :~.. ;.
Incubation of serum lyophilisa1;e to reduce turbidity in .
reconstituted solution : 2 batches of Lp(a)-containing control serum prepared according~to the method of AT~B-361,135 and .
having a sucrose content of 15 ~ were lyophilized and :.
subsequently incubated at 37C in a moist atmosphere.
The increase in the water content of the lyophilisate .
as well as the decrease~of turbidity in the reconstituted solution were determined after 1, ~, 3 and 4 weeks. -:: ~ ..
Table 4 Influence of incubation time on water content in serum lyophilisate and on turbidity of reconstituted solution Batch 1 Batch 2 Incubation Turbidity of Water Turbidity of Water time (d) rec. sol. content rec. sol. content ~ -~
(LSU) (~) (LSU) (~) 0 88 1.2 270 1.7 7 49 1.3 228 2.4 14 3B 1.4 181 j 3.2 28 14 2.5 40 _3,6 ,,'',.- :".~, ....
The lyophilization of aqueous solutions is a common method of producing preparations that are stable during long-term storage. In doing so, the water is removed from the frozen solution by sublimation under vacuum. The water content of storage-stable lyophilisates, in general, amounts to less than 1 %
(w/w). Yet, rendering aqueous solutions storabLe by lyophilization is suitable only if the physico-chemical properties of the ingredients will not be substantially changed by the removal of water~
The lyophilizatlon of lipoprotein-containing plasma or serum constitutes a great problem, since, according to the experience, turbid solutions are obtained upon reconstitution. Lipoproteins may be denatured by lyophilization such that their determlnation by means of electrophoretic or immunologic or optical methods is difficult. A common ; ~ -measure for the turbidity of a solution is LSU (light scattering unit). In general, a solution is considered turbid if it has a measured signal of more than 70 LSU
when measured in the nephelometer at 340 nm and a path length of 1 cm (reference: water).
From the document Clin. Chem. (1990), 36, 366-369, ~. ,, . ' .
`:~
it is apparent that lyophilized serum is not suitable as a reference for determining lipoproteins because of the denaturation of lipoproteins. This holds, in particular, for apo-B lipoproteins, whos~ structure is ., .~
changed during dehydration and rehydration in such a manner that their solubility is reduced, which results in the turbidity of the serum.
Attempts to prevent the formation of turbidity by the addition of stabilizers, such as, e.g., sucrose, to the starting material (serum) have been described in AT B-361,135 and in US-A-4,701,417. Accordingly,, the turbidity, expressed by the LSU measured in - ~
nephelometry, can be inadmissikly high such that these -preparations cannot be applied as references for nephelometric determination. A solution is considered suitable for use in diagnostic methods i it has a maximum turbidity of 70 LSU.
A special method of lyophilization is described in US-A-3,928,566 and US-A-3,932,943. Frozen droplets are formed in the serum or plasma by a sprayin~ procedure, thereby causing an increase in turbidity upon reconstitution of less than 20 ~. However, this method involves high expenditures.
It is the object of the invention to avoid the drawbacks of lipoprotein-containing lyophilisates ~ ~
mentioned above and to provide a lyophilized co~trol or ~ ~ `
reference plasma or control or reference serum for `~
diagnostic purposes, which is capable o~ being reconstitued to a clear solution and can be produced by a simple method.
In accordance with the invention, this object is achieved b~ a lyophilized control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular apo-B-containing lipoproteins, which is characterized in tha$ it has a water content ranging between 1 and 10 ~, preferably between 1.5 and 5 ~, and is reconstitutable to a clear solution exhibiting a maximum turbidity of 70 LSU.
A particularly preferred range of the water content of the lyophilisate is between 2 and 5 %.
It has been shown that, due to a given residual moistura in the lyophilisate, reconstitution to a clear solution is possible. The water content in the lyophilisate is indirectly related to the increase in turbidity by lyophilization. The higher the water content of the hydrated lyophilisate, the low~r the increase in turbidity. A residual moisture of between 1 and 10 ~ (w/w) is suitable primarily because the storage stability may be jeopardized at a higher water content. ~;~
It has also been found that the increase in -~
turbidity due to lyophilization and reconstitution is ~ -r ev~n larger if already the starting material is relatively turbid. Hence, it must be taken care that it is depar~ed from a suitable starting material in order to measu~e not more than 70 LSU in the reconstituted -::
solution. Values of above 70 LSU correspond to a turbidity that renders the solution practically unsuitable for use as a reference or control in nephelometric determinations. Values of up to 50 LSU
are particularly suitable.
The lyophilized control or reference plasma, or ; -control or reference serum according to the invention, ;~
furthermore, may contain non-reducing sugars, such as sucrose, for stabilization. A content of sucrose that ~ `
does not exceed 25 ~, preferably ranging between 12 and 20 %, is advantageous. ;~
The plasma or serum according to the invention is sufficient'y stable for long-term storage. This was not to be foreseen, because the water content was generally reduced in a lyophilisate as far as possible in order ; ;~
to improve the s~orage stability.
The invention, furthermore, relates to a method of preparing a lyophilized control or reference plasma or control or reference serum which is characterized in that lipoprotein-containing plasma or serum optionally is admixed with a non-reducing sugar, preferably sucrose, and is lyophilized until reaching a water content of between 1 and 10 ~
It has been shown that a turbidity of more than 70 LSU in the reconstituted solution can be prevented if -the lyophilisate is incubated in the moist sta~e or in a moist atmosphere. In doing so, the lyophilisate reabsorbs moisture in a reversible manner, which, in turn, results in a clear reconstituted solution. Thus, lyophilisa~es that would yield turbid and hence unusable solutions upon con~entional reconstitution (addition of water) are capable of being reconstituted to solutions ha~ing a turbidity of less than 70 LSU iand thus being usable in diagnostics.
A preferred method of preparing a lyophilized control or reference plasma or control or reference serum, therefore, is characterized in that a lyophilized lipoprotein-containing plasma or serum is incubated in a moist atmosphere. ~ ~;
Another embodiment of the method of preparing a lyophilized control or reference plasma or control or reference serum consists in that a lipoprotein-containing plasma or serum optionally is admixed with non-reducing sugars, preferably sucrose, and is lyophilized and the lyophilisate is adjusted to a water ontent of between 1 and 10 ~ by incubation in a moist atmosphere.
; Thus, con~rol or reference plasmas or control or reference sera that exhibit a maximum turbidity of 70 LSU even can be prepared from lyophilisates having low watericontents, by incubation in a moist atmosphere.
~`~ A method of preparing a control or reference plasma or control or reference serum for diagnostic purposes containing lipoproteins, in particular, apo-B~
containing lipoproteins is characterized in that a -lyophilized control or reference plasma or control or - 5 - ;
y i ~ :
reference serum according to the present invention is : :
reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU.
A particularly preferred embodiment of the method of preparil~g a control or reference plasma or control or reference serum for diagnostic purposes containing ~ ~
lipoproteins, in particular, apo-B-containing ~ :
lipoproteins consists in that a lyophilized lipoprotein-containin~ plasma or serum is adjusted to a water content of between 1 and 10 ~ by incubation in a moist atmosphere and subsequently is reconstituted to a clear solution exhibiting a maximum turbidity of 70 ~ -.
LSU. ~ ;
Incubation in a moist atmosphere is effected at appropriate temperatures ranging from 4 to 50C, preferably from 30 to 40C, in particular, at 37C, ~: ;
until the turbidity of the reconstituted solution;~
corresponds to a maximum value of 70 LSU. The duration of Lncubation is a funation of the temperature and may last from 1 to 50 da~s, pre~erably from 7 to 28 days.
: The addition of non-reducin~ sugars, in particular : suorose, may be effected prior to lyophilization~
' The invention wlll be explained in more detail by :~ the following examples.
ExampIe 1 Preparation of plasma lyophilisates having different water contents --~
: Rethrombinized fresh plasma samples were mixed : ;: .''.', ~,`
; - 6 -~ .
with 15 % sucrose and examined for turbidity. After this, samples with 20 LSU were lyophilized to a residual moisture of 0.5, 1.1, 3.8 and 8.5 % (w/w) water. For comparison, plasma samples with 48 LSU were lyophilized to a residual moisture of l.1 and 2.8 ~
(w/w). The water content was measured according to Karl Fischer's technique.
The lyophilisates were reconstituted with water and diluted until reaching their starting volumes. The turbidities of the reconstituted solutions were measured by an IMMUN0 video nephPlometer (IMMUN0 AG).
Table 1 indicates that the turbidity of the solution of the lyophilisate having a water content of 0.5 % (w/w) is inadmissibly high. Likewise, it is ;
apparent that plasma exhibiting an elevated self~
turbidity brings about a still admissible turbidity in ~-the reconstituted solution only at an elevated water content of the lyophilisate. There is a linear relationship between the logarithm of the water content and the logarithm of the ratio of the turbidity exhibited by the reconstituted solution to that exhibited by the starting plasma (r = -0.833.
; ' ;'' . '' '~ - ' Table 1 Influence of water content in plasma lyophilisate on turbidity in reconstituted solution Water content inTurbidity in Turbidity in lyophilisatestarting plasmareconstituted (~, w/w) (LSU) solution ~LSU) 0.5 20 100 1.1 20 70 ~:
3.8 ZO ~6 8.5 20 19 1.1 48 ~00 2.8 48 69 Example 2 ~;
Assessment of the storage stability of a control serum as a function of its residual moisture Lp(a)-containing control sera prepared accordi~g to the method of AT-B-361,135 and having a content of sucrose (15 ~) and differeht contents of water were ncubated at 37C for 14, 21 and 28 days. According to : experience, 28 days at 37C correspond to a storage , stability of 2 years at +4C. To assess its stability, Lp(a) (lipoprotein a) was quantitatively determined and the quotient QLp(a) was cal~ulated.
" ,, : Lp(a)-contentn days 37C
QLP ~ a) =-~ ~~
Lp(a)-co~entO days 37C ~ ;
.~
Table 2 Storage stability of control serum having varying water contents ' Water content QLP( a) after an incubation time of (%) 0 d 14 d 21 d 28 d .
3.0 1.00 1.00 1~00 1~04 : ~ ~ :
1.5 1.00 0.94 1.06 l~00 1.0 l.00 1,07 1.05 1.04 0.5 l.00 0.94 0.91 0.94 For the period of time investigated, the storage - `~
stability of the Lp(a)-containing serum is independent of its residual moisture. :
Example 3 Preparation of serum lyophilisates having varying sucrose contents Lp(a)-containing control serum prepared according to the method of AT-B-361,135 was mixed with varying portions of sucrose (S %, 10 %, 15 %), frozen and lyophilized in a manner that the water content varied in the end product. -, ~.
;,~
~:
Table 3 Influence of sucrose content in serum lyophilisate ~ :
on turbidity of reconstituted solution Turbidity (LSU) in reconstituted solution of serum lyophilisate having a sucrose content of Water content (%) 5 % 10 % 15 % ; . `~
sucrose 8.0 - 8.4 2~0 LSU 57 LSU n.d.*
2,9 - 4,2 195 LSU 95 LSU 41 LSU
1,0 - 1.2 _ 210 LSU 194 LSU , 73 LSU ~
*) .... not determined ~ .
Example 4 - :~.. ;.
Incubation of serum lyophilisa1;e to reduce turbidity in .
reconstituted solution : 2 batches of Lp(a)-containing control serum prepared according~to the method of AT~B-361,135 and .
having a sucrose content of 15 ~ were lyophilized and :.
subsequently incubated at 37C in a moist atmosphere.
The increase in the water content of the lyophilisate .
as well as the decrease~of turbidity in the reconstituted solution were determined after 1, ~, 3 and 4 weeks. -:: ~ ..
Table 4 Influence of incubation time on water content in serum lyophilisate and on turbidity of reconstituted solution Batch 1 Batch 2 Incubation Turbidity of Water Turbidity of Water time (d) rec. sol. content rec. sol. content ~ -~
(LSU) (~) (LSU) (~) 0 88 1.2 270 1.7 7 49 1.3 228 2.4 14 3B 1.4 181 j 3.2 28 14 2.5 40 _3,6 ,,'',.- :".~, ....
Claims (28)
1. A lyophilized control or reference plasma or control or reference serum for diagnostic purposes with a content of lipoproteins, which comprises a water content ranging between 1 and 10 % and is capable of being reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU.
2. A lyophilized control or reference plasma or control or reference serum as set forth in claim 1, wherein said lipoproteins are apo-B-containing lipoproteins.
3. A lyophilized control or reference plasma or control or reference serum as set forth in claim 1, which comprises a water content ranging between 1.5 and 5 %.
4. A lyophilized control or reference plasma or control or reference serum as set forth in claim 1, further comprising a non-reducing sugar.
5. A lyophilized control or reference plasma or control or reference serum as set forth in claim 4, wherein said non-reducing sugar is sucrose at a concentration of from 12 to 20 %.
6. A method of preparing a lyophilized control or reference plasma or control or reference serum for diagnostic purposes having a content of lipoproteins, a water content of between 1 and 10 % and capable of being reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU, which method comprises providing a lipoprotein-containing plasma or serum and lyophilizing said lipoprotein-containing plasma or serum until a lyophilized lipoprotein-containing plasma or serum having a water content of between 1 and 10 %
is obtained.
is obtained.
7. A method as set forth in claim 6, wherein said lipoproteins are apo-B-containing lipoproteins.
8. A method as set forth in claim 6, further comprising adding a non-reducing sugar to said lipoprotein-containing plasma or serum.
9. A method as set forth in claim 8, wherein said non-reducing sugar is sucrose.
10. A method of preparing a lyophilized control or reference plasma or control or reference serum for diagnostic purposes having a content of lipoproteins, a water content of between 1 and 10 % and capable of being reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU, which method comprises providing a lyophilized lipoprotein-containing plasma or serum and incubating said lyophilized lipoprotein-containing plasma or serum in a moist atmosphere to adjust a water content of between 1 and 10 % in said lyophilisate.
11. A method as set forth in claim 10, wherein said lipoproteins are apo-B-containing lipoproteins.
12. A method as set forth in claim 10, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 5 and 50°C and for a period of time ranging between 1 and 50 days.
13. A method as set forth in claim 10, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 30 and 40°C and for a period of time ranging between 7 and 28 days.
14. A method of preparing a lyophilized control or reference plasma or control or reference serum for diagnostic purposes having a content of lipoproteins, a water content of between 1 and 10 % and capable of being reconstituted to a clear solution exhibiting a maximum turbidity of 70 LSU, which method comprises providing a lipoprotein-containing plasma or serum, lyophilizing said lipoprotein-containing plasma or serum so as to obtain a lyophilisate and incubating said lyophilisate in a moist atmosphere so as to adjust a water content of between 1 and 10 % in said lyophilisate.
15. A method as set forth in claim 14, wherein said lipoproteins are apo-B-containing lipoproteins.
16. A method as set forth in claim 14, further comprising adding a non-reducing sugar to said lipoprotein-containing plasma or serum.
17. A method as set forth in claim 16, wherein said non-reducing sugar is sucrose.
18. A method as set forth in claim 14, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 4 and 50°C and for a period of time ranging between 1 and 50 days.
19. A method as set forth in claim 14, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 30 and 40°C and for a period of time ranging between 7 and 28 days.
20. A method of preparing a control or reference plasma or control or reference serum for diagnostic purposes having a content of lipoproteins and exhibiting a maximum turbidity of 70 LSU, which method comprises providing a lyophilized control or reference plasma or control or reference serum having a water content ranging between 1 and 10 % and reconstituting said lyophilized control or reference plasma or control or reference serum to a clear solution.
21. A method as set forth in claim 20, wherein said lipoproteins are apo-B-containing lipoproteins.
22. A method as set forth in claim 20, wherein said lyophilized control or reference plasma or control or reference serum further comprises a non-reducing sugar.
23. A method as set forth in claim 22, wherein said non-reducing sugar is sucrose at a concentration of from 12 to 20 %.
24. A method as set forth in claim 20, wherein said lyophilized control or reference plasma or control or reference serum comprises a water content ranging between 1.5 and 5 %.
25. A method of preparing a control or reference plasma or control or reference serum for diagnostic purposes having a content of lipoproteins and exhibiting a maximum turbidity of 70 LSU, which method comprises providing a lyophilized lipoprotein-containing control or reference plasma or control or reference serum, adjusting said lyophilized control or reference plasma or control or reference serum to a water content of between 1 and 10 % by incubation in a moist atmosphere and subsequently reconstituting said lyophilized control or reference plasma or control or reference serum to a clear solution.
26. A method as set forth in claim 25, wherein said lipoproteins are apo-B-containing lipoproteins.
27. A method as set forth in claim 25, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 4 and 50°C and for a period of time ranging between 1 and 50 days.
28. A method as set forth in claim 25, wherein said incubating in a moist atmosphere is effected at a temperature ranging between 30 and 40°C and for a period of time ranging between 7 and 28 days.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AT55393A AT399056B (en) | 1993-03-19 | 1993-03-19 | LYOPHILIZED CONTROL OR REFERENCE PLASMA OR CONTROL OR REFERENCE SERUM FOR DIAGNOSTIC PURPOSES |
| ATA553/93 | 1993-03-19 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2119096A1 true CA2119096A1 (en) | 1994-09-20 |
Family
ID=3493660
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2119096 Abandoned CA2119096A1 (en) | 1993-03-19 | 1994-03-15 | Lyophilized control or reference plasma or control or reference serum for diagnostic purposes |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP0617289A3 (en) |
| JP (1) | JPH06300758A (en) |
| AT (1) | AT399056B (en) |
| CA (1) | CA2119096A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE358683T1 (en) * | 1999-06-02 | 2007-04-15 | Kyowa Medex Co Ltd | STABILIZED DENatured LIPOPROTEIN AND METHOD FOR THE PRODUCTION THEREOF |
| JP4534511B2 (en) * | 2004-02-16 | 2010-09-01 | 東ソー株式会社 | Lyophilized samples containing lipoproteins |
| CN114279778A (en) * | 2021-12-04 | 2022-04-05 | 南京岚煜生物科技有限公司 | Preparation method of composite high-value reference product |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4121905A (en) * | 1977-08-22 | 1978-10-24 | Jonas Maurukas | Process for preparing biological compositions for use as reference controls in diagnostic analyses |
| AT361135B (en) * | 1979-06-27 | 1981-02-25 | Immuno Ag | METHOD FOR PRESERVING THE ELECTRO-PHORETIC PROPERTIES OF LIPOPROTEINS |
| US4659699A (en) * | 1983-08-22 | 1987-04-21 | Cetus-Ben Venue Therapeutics | Process for freeze drying cyclophosphamide |
| DE3445010A1 (en) * | 1984-12-10 | 1986-06-19 | Boehringer Mannheim Gmbh | CONTROL OR OAK SERUM FOR LIPID DIAGNOSTICS |
-
1993
- 1993-03-19 AT AT55393A patent/AT399056B/en not_active IP Right Cessation
-
1994
- 1994-03-09 EP EP94890054A patent/EP0617289A3/en not_active Withdrawn
- 1994-03-15 CA CA 2119096 patent/CA2119096A1/en not_active Abandoned
- 1994-03-18 JP JP4852694A patent/JPH06300758A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| AT399056B (en) | 1995-03-27 |
| EP0617289A3 (en) | 1995-09-06 |
| EP0617289A2 (en) | 1994-09-28 |
| JPH06300758A (en) | 1994-10-28 |
| ATA55393A (en) | 1994-07-15 |
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