In Vivo Recovery of Factor VIII: A Comparison of One-Stage and Two-Stage Assay Methods

1979 ◽  
Vol 42 (04) ◽  
pp. 1230-1239 ◽  
Author(s):  
I M Nilsson ◽  
T B L Kirkwood ◽  
T W Barrowcliffe
Keyword(s):  
Factor Viii ◽  
Half Life ◽  
In Vitro Assays ◽  
Two Stage ◽  
One Stage ◽  
Significant Difference ◽  
Assay Methods ◽  
The One ◽  
Fraction I

SummaryThe recovery and half-life of VIII: C in the plasma of severely haemophilic patients was measured by one-stage and two-stage assays after injection of two Factor VIII concentrates (Hemofil, Hyland and Fraction I-O, Kabi). Plasma volumes were measured with an Evans� Blue technique, and both concentrates and post-infusion samples were measured against the same plasma standard.There was a highly significant difference in recoveries estimated by the two assay methods. The one-stage assays gave the most consistent results, in that the average recovery was 100%, whereas the two-stage assays gave only about 80% of the value expected from in vitro assays. There was no difference in recoveries between the two concentrates.The two-stage assays gave a slightly shorter half-life than the one-stage assays, and the half-life of Hemofil was also shorter than that of Fraction I-O.

Decreased Potency Of Factor VIII Concentrates

1981 ◽  
Author(s):  
C K Kasper
Keyword(s):  
Factor Viii ◽  
In Vitro Assays ◽  
Assay Method ◽  
Two Stage ◽  
One Stage ◽  
Plasma Factor ◽  
Factor Viii Concentrates ◽  
The One

Plasma factor VIII recoveries after infusions of factor VIII concentrates into patients with classic hemophilia have been measured in this laboratory for 14 years. Recently, we observed a decline in the in vivo recovery of factor VIII per factor VIII unit infused. In 1980, plasma factor VIII levels were measured by a one-stage APTT-based assay before and 10 min after 150 infusions of 46 lots of 3 brands of factor VIII concentrate produced in the U.S.A. Our pooled normal plasma reference was calibrated against WHO International Standard 2 and results expressed in International factor VIII units. Observed in vivo factor VIII recovery was compared to the value expected from calculations based on the unitage stated on the label. The ratio of observed/expected recovery averaged 56% per lot for brand A, 60% per lot for brand B, and 103% per lot for brand C. In vitro assays were performed on 22 lots on 36 occasions, and the ratio of observed/labelled units average 46% per lot for brand A, 53% for brand B and 75% for brand C. The two-stage factor VIII assay method of Pool and Robinson was also used to assay plasma samples from 18 infusions, and results averaged 135% of the one-stage values for infusions of brand A, 160% for brand B, and 109% for brand C. (Brand A is assayed by the manufacturer by a two-stage method, brands B and C by one-stage methods.)Decreased clinical efficacy was observed when postinfusion plasma factor VIII levels were lower than customary. The decline in potency of brands A and B has necessitated more frequent assay of patients and use of larger amounts of concentrate, with greatly-increased expense. Investigation of the effect of different assay methods and different factor VIII standards and references on the apparent factor VIII content of concentrates has begun.

scholarly journals Assessing the Performance of Extended Half-Life Coagulation Factor VIII, FC Fusion Protein by Using Chromogenic and One-Stage Assays in Saudi Hemophilia A Patients

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Tarek M. Owaidah ◽  
Hazzaa A. Alzahrani ◽  
Nouf S. Al-Numair ◽  
Abdulmjeed O. Alnosair ◽  
Amelita M. Aguilos ◽  
...  
Keyword(s):  
Factor Viii ◽  
Half Life ◽  
Hemophilia A ◽  
Coagulation Factor ◽  
Time Interval ◽  
One Stage ◽  
Chromogenic Assay ◽  
Significant Difference ◽  
Mean Differences ◽  
The One

Background. The one-stage assay is the most common method to measure factor VIII activity (FVIII : C) in hemophilia A patients. The chromogenic assay is another two-stage test involving purified coagulation factors followed by factor Xa-specific chromogenic substrate. Aim. This study aimed to assess the discrepancy and correlation between the chromogenic and one-stage assays in measuring FVIII : C levels in hemophilia patients receiving Extended Half-Life Elocta® as a recombinant extended half-life coagulation factor. Methods. We performed a study comparing the measurements of FVIII : C levels by the chromogenic versus the one-stage assays at different drug levels. Data of FVIII : C levels, dosage, and the time interval from administration to measurement were retrieved from the hospital records. The correlation, mean differences, and discrepancy between the two assays were calculated. The linear regression analysis was used to predict the time interval till reaching 1% FVIII : C. Results. Fourteen patients with 56 samples were included in the study. Of them, 13 patients were receiving Elocta® as a prophylactic, while one was receiving Elocta® on demand. One-third of these samples showed a discrepancy between the chromogenic and one-stage assays. The two assays were well correlated. Mean differences were significant at the individual and the time interval level. The time since the last Elocta® injection could significantly predict FVIII : C levels (β = 0.366, P<0.001). Conclusion. Our findings suggested a significant difference between both methods; the FVIII : C levels measured by the one-stage assay were less than those estimated by the chromogenic assay. However, the measurements of FVIII levels by the two assays were well correlated but discrepant in one-third of the samples. The levels of FVIII : C reach 1% after 5.4 days since the last Elocta® administration.

In Vivo Recovery and Survival of Monoclonal-Antibody-Purified Factor VIII Concentrates

1991 ◽  
Vol 66 (06) ◽  
pp. 730-733 ◽  
Author(s):  
Carol K Kasper ◽  
Hugh C Kim ◽  
Edward D Gomperts ◽  
Kenneth J Smith ◽  
Phyllis M Salzman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Factor Viii ◽  
In Vitro Assays ◽  
Two Stage ◽  
Double Blind ◽  
One Stage ◽  
Factor Viii Concentrates ◽  
In Vivo Recovery

SummaryIn response to reports of discrepant in vitro assays of high-purity concentrates, a double-blind crossover study of in vivo recovery and half-life of two brands of monoclonal-antibody-purified factor VIII concentrates (Monoclate and Hemofil-M) was performed in 23 patients with hemophilia A. In vivo recoveries were close to values predicted from the labelled unitage when plasma samples were assayed by a one-stage method. When a two-stage assay was used, lower recoveries were calculated and the recovery with Hemofil-M was slightly but significantly lower than that with Monoclate. The concentrates were re-assayed in vitro by the two-stage method. Monoclate (which is assayed by the manufacturer using a two-stage method) contained 97% of the labelled potency and Hemofil-M (which is assayed by the manufacturer using a one-stage method) contained 81% of the labelled potency. Differences in in vitro and in vivo assay methods contribute to disparities between expected and observed factor VIII recovery. Clearance of Hemofil-M was significantly faster than that of Monoclate, but volume of distribution at the steady state, mean residence time, and plasma half-disappearance times of the two concentrates were not significantly different.

Studies Of Factor VIII Inhibitor Bypassing Activity (FEIBA)

1981 ◽  
Author(s):  
T W Barrowcliffe ◽  
E Gray ◽  
G Kemball-Cook
Keyword(s):  
Factor Viii ◽  
Factor Xa ◽  
Factor Ix ◽  
Procoagulant Activity ◽  
Two Stage ◽  
Human Antibodies ◽  
One Stage ◽  
Viii And Ix ◽  
The One

The activities of an activated Factor IX concentrate (FEIBA, Immuno AG) were studied by two in vitro assays: a one-stage method using VUI-deficient plasma as substrate, and a two-stage assay based on the thrombin generation test. The nature of the active principle was explored by measuring the reduction in activity when FEIBA was incubated with specific antibodies.Incubation of FEIBA with human antibodies to Factor VIII reduced its activity by about 30% in the one-stage assay, and about 50% in the two-stage assay, suggesting that FEIBA contains Factor VIII procoagulant activity. Inactivation of the Factor VIII in FEIBA was somewhat slower than that of normal Factor VIII, indicating partial protection from inhibition. Human antibodies to Factor IX inhibited the one stage activity by about 30%, and incubation with both antibodies also produced a 30% reduction in activity. The remaining procoagulant activity decayed only slowly when incubated with non-inhibitor plasma. In contrast, purified human Factor Xa lost activity rapidly on incubation in normal plasma, as did a purified fraction from a Factor IX concentrate, which had high activity in the one-stage assay.These results suggest that the in vitro activity of FEIBA is due to at least two components. One component appears to be dependent on both Factors VIII and IX and may be a complex of VIII and IXa. The other component acts later than Factors VIII and IX in the coagulation cascade but, unlike purified Factor Xa, is relatively resistant to inactivation by plasma inhibitors such as antithrombin III. FEIBA was also found to contain phospholipid, and it may be that the phospholipid protects both the Factor VIII and activated enzymes from their inhibitors.

Factor VIII: C (FVIII: C) Recovery and Half-Life After Infusion of Steam-Treated High Purity Factor VIII Concentrate in Severe Hemophilia A - Comparison of One-Stage Assay, Two-Stage Assay and a Chromogenic Substrate Assay

1986 ◽  
Vol 56 (03) ◽  
pp. 353-359 ◽  
Author(s):  
Peter Hellstern ◽  
Ralf Kiehl ◽  
Chieko Miyashita ◽  
Holger Schwerdt ◽  
Gerhard von Blohn ◽  
...  
Keyword(s):  
Factor Viii ◽  
High Purity ◽  
Plasma Levels ◽  
Half Life ◽  
Hemophilia A ◽  
Least Square ◽  
Chromogenic Substrate ◽  
Two Stage ◽  
One Stage ◽  
The One

SummaryFactor VIII :C recovery and half-life was measured in 16 hemophilia A patients under comprehensively standardized conditions. Each patient received the same lot of a steam-treated high purity FVIII concentrate at a dose of 19-33 U/kg body weight. A comparison was made between the one-stage assay, the two-stage assay and a chromogenic substrate test for FVIII :C determination using a FXa-sensitive chromogenic substrate. Factor VIII :C potency of the administered FVIII concentrate was measured using calibration curves derived from a concentrate standard and FVIII: C plasma levels were read from calibration curves derived from a plasma standard. The chromogenic assay showed a good reproducibility at FVIII: C levels between 0.015 and 0.50 U/ml. The FVIII :C recoveries calculated from the results of the one-stage assay, the two-stage assay and the chromogenic substrate test were 109 ± 20, 92 ± 14 and 81 ± 11% (x ± SD), respectively. The elimination half-lives of FVIII :C were calculated by non-linear least square analysis using a modified computerized Gauss-Newton algorithm. The half-lives calculated from the FVIII: C plasma levels measured by the one-stage assay, the two-stage assay and the chromogenic test were 23.8 ± 6.4, 22.2 ± 5.7 and 17.1 ± 4.8 h (x ± SD), respectively. No previous study has reported such long half-life values. Our findings indicate that measurements of recoveries and half-lives by the chromogenic FVIII :C assay and by computerized nonlinear least square analysis allow the possibility of individualized FVIII replacement therapy.

Standardization of Factor VIII – IV. Establishment of the 3rd International Standard for Factor VIII: C Concentrate

1983 ◽  
Vol 50 (03) ◽  
pp. 697-702 ◽  
Author(s):  
T W Barrowcliffe ◽  
A D Curtis ◽  
D P Thomas
Keyword(s):  
Factor Viii ◽  
High Purity ◽  
Collaborative Study ◽  
World Health ◽  
Current Standard ◽  
Two Stage ◽  
International Standard ◽  
One Stage ◽  
The One ◽  
Health Organization

SummaryAn international collaborative study was carried out to establish a replacement for the current (2nd) international standard for Factor VIII: C, concentrate. Twenty-six laboratories took part, of which 17 performed one-stage assays, three performed two-stage assays and six used both methods. The proposed new standard, an intermediate purity concentrate, was assayed against the current standard, against a high-purity concentrate and against an International Reference Plasma, coded 80/511, previously calibrated against fresh normal plasma.Assays of the proposed new standard against the current standard gave a mean potency of 3.89 iu/ampoule, with good agreement between laboratories and between one-stage and two- stage assays. There was also no difference between assay methods in the comparison of high-purity and intermediate purity concentrates. In the comparison of the proposed standard with the plasma reference preparation, the overall mean potency was 4.03 iu/ampoule, but there were substantial differences between laboratories, and the two-stage method gave significantly higher results than the one stage method. Of the technical variables in the one-stage method, only the activation time with one reagent appeared to have any influence on the results of this comparison of concentrate against plasma.Accelerated degradation studies showed that the proposed standard is very stable. With the agreement of the participants, the material, in ampoules coded 80/556, has been established by the World Health Organization as the 3rd International Standard for Factor VIII :C, Concentrate, with an assigned potency of 3.9 iu/ampoule.

scholarly journals Identification of Sources of Inter-Laboratory Variation in Factor VIII Assay

1977 ◽  
Author(s):  
T.B.L. Kirkwood ◽  
C.R. Rizza ◽  
T.J. Snape ◽  
I. Rhymes ◽  
D.E.G. Austen
Keyword(s):  
Factor Viii ◽  
Systematic Difference ◽  
Normal System ◽  
Two Stage ◽  
Initial Dilution ◽  
One Stage ◽  
Collaborative Studies ◽  
Freeze Dried ◽  
Sources Of Variation ◽  
The One

A repeated finding of national and international collaborative studies of standard Factor VIII preparations has been that systematic differences exist between laboratories in their measurement of the relative activities of the same pairs of Factor VIII preparations.A workshop meeting was held at the Oxford Haemophilia Centre (England) during 23rd-26th November 1976 to investigate which of the possible sources of variation between laboratories were responsible. Participants from 16 British laboratories (9 one-stage, 7 two-stage) performed a total of 273 assays using three freeze-dried preparations of differing purity (a plasma, an intermediate and a high purity concentrate). The results of assays with each participant using their normal system established that, if the participants were a representative cross-section, approximately one-third of one-stage laboratories would show a systematic difference from the overall mean of at least 16%, with a similar figure for the two-stage laboratories of 9%. Various features of the assay systems were then modified in a controlled series of experiments. The results showed conclusively that i) differences between reagents accounted for most of the variation between laboratories and, ii) the two-stage assays were, on average, detecting relatively more activity in the more purified preparations than the one-stage assays. The results also suggested that the use of buffer as opposed to haemophilic plasma for the initial dilution of concentrates did not affect the assay results.

scholarly journals Plasma Transfusions in Hemophilia

Blood ◽  
1952 ◽  
Vol 7 (7) ◽  
pp. 710-720 ◽  
Author(s):  
S. VAN CREVELD ◽  
M. M. P. PAULSSEN
Keyword(s):  
Two Stage ◽  
Coagulation Time ◽  
Prothrombin Activity ◽  
Lasting Effect ◽  
One Stage ◽  
The One ◽  
Rapid Appearance

Abstract Transfusions of heparinized plasma have a greater and more lasting effect on the coagulation time of hemophiliacs than transfusions of citrated plasma. Both in vitro and in vivo, heparinized plasma causes in hemophiliacs a far greater consumption of prothrombin as determined with the two-stage method than citrated plasma. In using the one-stage method no important differences in prothrombin activity are found after transfusions of heparinized and of citrated plasma respectively. This fact was thought to be connected with the more or less rapid appearance of an accelerator. Its a hemophiliac with a circulating anticoagulant, transfusions of heparinized plasma were unable to shorten the coagulation time to any important degree, nor did these transfusions cause an important decrease of serum prothrombin as determined by the two-stage method.

Surgical Management of Simultaneous Anterior Cruciate Ligament and Patellar Tendon Ruptures: A Systematic Review

2017 ◽  
Vol 31 (09) ◽  
pp. 875-883 ◽  
Author(s):  
Carlos Meheux ◽  
Robert Jack ◽  
Patrick McCulloch ◽  
David Lintner ◽  
Joshua Harris
Keyword(s):  
Systematic Review ◽  
Anterior Cruciate Ligament ◽  
Acl Reconstruction ◽  
Cruciate Ligament ◽  
Activity Level ◽  
Two Stage ◽  
One Stage ◽  
Significant Difference ◽  
Anterior Cruciate ◽  
The One

AbstractThis study performs a systematic review to determine (1) if a significant difference exists in return to preinjury activity level between one- and two-stage treatment of combined anterior cruciate ligament (ACL) and patellar tendon (PT) tears; and (2) if a significant difference exists in the number of postoperative complications between the two differing surgical treatment approaches. A systematic review was performed using Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines and registered on PROSPERO. MEDLINE, Cochrane Central Register of Controlled Trials, SCOPUS, and Sport Discus were searched for English language level I–IV evidence studies on either one- (simultaneous) or two-stage (sequential) surgical treatment of simultaneously sustained ipsilateral ACL and PT tears. The approach to initial evaluation, diagnosis, treatment, and outcomes were qualitatively analyzed. Methodological quality assessment of all included studies was completed using the Methodological Index for Non-randomized Studies (MINORS). The Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) tool was used to assess quality of evidence and provide strength of recommendation. Statistical analyses were done using Fischer's exact test. Eleven articles (18 patients; 83% males; mean age, 31.1 ± 10.1 years; mean follow-up, 2.2 ± 1.7 years; and mean MINORS 7.8/16) were analyzed. Eight patients had a one-stage procedure (primary PT repair and ACL reconstruction), and 10 patients underwent a two-stage procedure (primary PT repair first followed by ACL reconstruction) with mean 28 ± 45.7 weeks (5 weeks–3 years) between surgeries. The rate for return to preinjury activity level after surgery was not significantly different between one- (88%) and two-stage (100%) (p = 0.444). There was a significantly higher complication rate (p = 0.023) in the one-stage (stiffness, instability, and patella baja) versus two-stage surgery (no complications). There was no significant difference in return to preinjury activity level between one- and two-stage PT repair and ACL reconstruction. However, the one-stage combined surgery had a significantly higher complication rate compared with two-stage surgery. The level of evidence is IV.

scholarly journals Comparison of the Properties of Activated Carbons Produced in One-Stage and Two-Stage Processes

2018 ◽  
Vol 4 (3) ◽  
pp. 41 ◽  
Author(s):  
Davide Bergna ◽  
Toni Varila ◽  
Henrik Romar ◽  
Ulla Lassi
Keyword(s):  
Carbon Content ◽  
Activated Carbons ◽  
Total Carbon ◽  
Activation Process ◽  
Two Stage ◽  
One Stage ◽  
Surface Areas ◽  
Significant Difference ◽  
Stage Process ◽  
The One

Activated carbons (ACs) can be produced from biomass in a thermal process either in a direct carbonization-activation process or by first carbonizing the biomass and later activating the bio-chars into activated carbons. The properties of the ACs are dependent on the type of process used for production. In this study, the properties of activated carbons produced in one-stage and two-stage processes are considered. Activated carbons were produced by physical activation of two types of starting materials: bio chars produced from spruce and birch chips in a commercial carbonization plant and from the corresponding raw chips. The activated carbons produced were characterized regarding specific surfaces, pore volumes, and pore size distributions. The un-activated bio chars had varying surface areas, 190 and 140 m2 g−1 for birch and spruce, respectively, and pore volumes of 0.092 and 0.067 cm3 g−1, respectively. On the other hand, 530–617 and 647–679 m2 g−1 for activated bio chars from birch and spruce, respectively, and pore volumes 0.366–0.509 and 0.545–0.555 cm3 g−1, respectively, were obtained. According to the results obtained, two slightly different types of activated carbons are produced depending on whether a one-stage or a two-stage carbonization and activation process is used. The ACs produced in the one-stage process had higher specific surface areas (SSA), according to the BET-model (Brunauer–Emmett–Teller), compared to the ones produced in a two-stage process (761–940 m2 g−1 vs. 540–650 m2 g−1, respectively). In addition, total pore volumes were higher in ACs from the one-stage process, but development of micro-pores was greater compared to those of the two-stage process. This indicates that the process can have an influence on the ACs’ porosity. There was no significant difference in total carbon content in general between the one-stage and two-stage processes for spruce and birch samples, but some differences were seen between the starting materials. Especially in the one-stage procedure with 2 and 4 h steam activation, there was nearly a 10% difference in carbon content between the spruce and birch samples.

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